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ICAM-1/CD54 Antibodies

We offer ICAM-1/CD54 Antibodies for use in common research applications: Adhesion Blockade, Blocking/Neutralizing, ELISA, ELISA Capture (Matched Antibody Pair), ELISA Detection (Matched Antibody Pair), ELISA Standard (Matched Pair), Flow (Cell Surface), Flow Cytometry, Functional, Immunoassay, Immunocytochemistry, Immunocytochemistry/Immunofluorescence, Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, In vitro assay, Proximity Ligation Assay, Sandwich ELISA, Western Blot. Each ICAM-1/CD54 Antibody is fully covered by our Guarantee+, to give you complete peace of mind and the support when you need it. Our ICAM-1/CD54 Antibodies can be used in a variety of model species: Bovine, Canine, Human, Mouse, Rat, Sheep. Use the list below to choose the ICAM-1/CD54 Antibody which is most appropriate for your research; you can click on each one to view full technical details, images, references, reviews and related products. Choose from our ICAM-1/CD54 polyclonal antibodies and browse our ICAM-1/CD54 monoclonal antibody catalog.

Alternate Names for ICAM-1/CD54 Antibodies

anti-ICAM-1/CD54 antibody, anti-ICAM1 antibody, anti-BB2 antibody, anti-CD54 antibody, anti-CD54 antigen antibody, anti-cell surface glycoprotein P3.58 antibody, anti-human rhinovirus receptor antibody, anti-ICAM-1 antibody, anti-intercellular adhesion molecule 1 (CD54), human rhinovirus receptor antibody, anti-intercellular adhesion molecule 1 antibody, anti-Major group rhinovirus receptor antibody, anti-P3.58 antibody

38 Results for "icam-1-cd54" in Primary Antibodies

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AF796

ICAM-1/CD54 Antibody [Unconjugated]

(6 Reviews)

(88 Publications)

100 μg

AF796

ICAM-1/CD54 was detected in perfusion fixed frozen sections of mouse testis using Goat Anti-Mouse ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF796) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in Leydig cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Increased stress kinase signaling and JNK pathway-dependent cytokine and chemokine production by primary keratinocytes lacking BRAF and RAF1.(A) Reduced ERK phosphorylation and increased JNK/p38 activation in primary delta / delta ep2 keratinocytes stimulated with EGF and/or TNF alpha and IL1 beta for 15 min. (B) Increased cytokine and chemokine production in primary delta / delta ep2 keratinocytes treated with EGF, TNF alpha and IL1 beta for 24 hr. Cytokine and chemokine production was determined by multiplex analysis, except for TSLP which was quantified by ELISA. Data represent mean ± SEM of 3–5 biological replicates. (C–D) Cells were pretreated with D-JNKI1 inhibitors prior to stimulation with EGF, TNF alpha and IL1 beta for 15 min (C) or 24 hr (D). Data represent the mean ± SEM of technical replicates (n = 3). (E–F) Effect of shRNA-mediated Mlk3 silencing on ERK and JNK phosphorylation and ICAM1 expression (E; stimulation with EGF, TNF alpha and IL1 beta for 15 min) and on the expression of Ccl2 and Tslp mRNA (F; stimulation with EGF, TNF alpha and IL1 beta for 24 hr) by F/F2 and delta / delta ep2 keratinocytes. shRen, shRNA targeting Renilla, used as a control; sh1 and sh2, targeting Mlk3, binding sites nucleotide 2266–2285 and 2383–2402, respectively. The shRNAs were encoded by lentiviral vectors coexpressing GFP. GFP immunoblots are shown to confirm similar levels of infection in all samples. Data represent mean ± SEM of 4 biological replicates. Each keratinocyte culture represents a pool of three mice. Immunoblots are representative of three independent experiments. p1 = 0.041, p2 = 0.040, p3 = 1.89E-4, p4 = 0.018, p5 = 0.046, p6 = 0.020, p7 = 0.008, p8 = 0.016, p9 = 0.001, p10 = 0.018, p11 = 3.23E-4, p12 = 1.47E-4, p13 = 0.007, p14 = 0.03, p15 = 0.035, p16 = 0.023 and p17 = 0.046.DOI://dx.doi.org/10.7554/eLife.14012.018Compound knockdown (KD2) of BRAF and RAF1 induce the expression of inflammation markers by HaCat cells in a MLK3/JNK-dependent manner.(A) Reduced ERK and increased JNK/p38 activation in BRAF and RAF1 knockdown (KD2) HaCat cells stimulated with EGF, TNF alpha and IL1 beta for 15 min. (B) D-JNKI1 reduces ICAM1 and CCL2 (n = 4) expression in KD2 cells treated with TNF alpha . (C) MEKi induces ICAM1 and CCL2 (n = 3) expression in RAF1KD cells treated with TNF alpha . In (B–C), ICAM1 expression was measured after a 3 hr, CCL2 expression after a 24 hr treatment with TNF alpha . (D) Effect of MLK3 silencing on ERK and JNK phosphorylation in WT and KD2 cells stimulated as in (A). MLK3 was silenced using a pool of oligonucleotides targeting the following regions: 686–704; 1489–1507; 2122–2138; and 2348–2366. MLK3 KD cells stimulated as in (B–C) show a decrease in JNK activation, ICAM1 and CCL2 (n = 7) expression. Immunoblots are representative of three independent experiments. qPCR data represent mean ± SEM of three independent experiments run in duplicates (p1 = 4.62E-4, p2 = 0.013, p3 = 0.050, p4 = 8.60E-8, p5 = 0.050, p6 = 0.001, p7 = 0.001 and p8 = 0.012).DOI://dx.doi.org/10.7554/eLife.14012.019 Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/14012), licensed under a CC-BY license. Not internally tested by R&D Systems.

D-JNKI1 treatment rescues inflammation in delta / delta ep2 mice.Mice were treated with D-JNKI1 or TAT peptide (22 mg/kg i.p. at 10 days of age) and analyzed after 12 days (A) D-JNKI1 treatment prevents disease onset in delta / delta ep2 mice. Immunoblot of epidermal lysates showing the effect of D-JNKI1 on the phosphorylation and expression of the indicated proteins, quantified as in Figure 1F. ACTB is shown as a loading control. (B–D) Decreased eyelid inflammation, mast cells infiltration (B; TB+; quantified in the plot on the right), epidermal chemokine/cytokine levels (C) and activated T cells, B cells and dendritic cells in lymph nodes (D) in D-JNKI1-treated delta / delta ep2 mice. Scale bars, 25 µm. Data represent mean ± SEM (n = 3–5; p1 = 0.026, p2 = 0.042, p3 = 0.022, p4 = 0.048, p5 = 0.044, p6 = 0.020, p7 = 0.025, p8 = 0.018, p9 = 0.016, p10 = 0.014, p11 = 0.023, p12 = 0.011, p13 = 0.039, p14 = 0.049, p15 = 0.015, p16 = 0.003, p17 = 1.70E-4, p18 = 0.008, p19 = 0.008, p20 = 0.004, p21 = 0.003, p22 = 0.017, p23 = 0.026, p24 = 0.027, p25 = 0.005, p26 = 2.13E-6, p27 = 4.50E-8, p28 = 1.39E-5, p29 = 0.001, p30 = 0.001, p31 = 0.001, p32 = 0.023, p33 = 2.35E-4, p34 = 0.050, p35 = 0.002, p36 = 0.050 and p37 = 0.012).DOI://dx.doi.org/10.7554/eLife.14012.015K6 expression and epidermal chemokine and cytokine levels in D-JNKI1-treated mice.(A) K6 expression is indistinguishable in TAT or D-JNKI1 treated F/F2 and △/△ep2 littermates. Scale bars, 25 µm. (B) Inflammatory chemokines and cytokines in epidermal lysates of TAT or D-JNKI1-treated mice. Data represent mean ± SEM (n = 3–5; p1 = 0.005, p2 = 0.001, p3 = 0.043, p4 = 0.026, p5 = 0.032, p6 = 0.016 and p7 = 0.051).DOI://dx.doi.org/10.7554/eLife.14012.016The inflammatory phenotype of delta / delta ep2 mice is not rescued by MyD88, caspase 1/11, or TNF knockout.Macroscopic appearance, spleen and lymph node size and circulating blood cell analysis are shown for the indicated genotypes (A–C). (A) Representative pictures of 4 month old delta / delta ep2, delta / delta ep2 MyD88-/- and control animals. Plots on the right represent the ratio between total splenocytes or lymph node cell numbers and body weight (n = 3–4). (B) Representative pictures and hemogram of 4 month old delta / delta ep2, delta / delta ep2 caspase 1/11-/- and control animals (n = 5–6). (C) Representative pictures and hemogram of 4 month old delta / delta ep2, delta / delta ep2 TNF-/- and control animals (n = 4–5). The macroscopic appearance of at least ten mice per genotype was monitored. Data represent mean ± SEM. p1 = 0.041, p2 = 0.052, p3 = 0.024, p4 = 0.023, p5 = 0.026, p6 = 0.034, p7 = 0.023, p8 = 0.007, p9 = 0.031, p10 = 0.011, and p11 = 0.034.DOI://dx.doi.org/10.7554/eLife.14012.017 Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/14012), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Species Glossary

Reactivity: Mu

Applications:

, ,

Applications: , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF796

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF796-SP

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BBA3

ICAM-1/CD54 Antibody (BBIG-I1 (11C81)) [Unconjugated]

(4 Reviews)

(81 Publications)

100 μg

BBA3

Western blot shows lysates of C32. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ICAM‑1/CD54 Monoclonal Antibody (Catalog # BBA3) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for ICAM‑1/CD54 at approximately 85 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Western Blot Buffer Group 1.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BBA9
Fluorescein	BBA20

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	BBA20

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NBP2-22541

ICAM-1/CD54 Antibody (1A29)

(1 Review)

(17 Publications)

0.1 mg

NBP2-22541

Flow Cytometry: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Analysis of CD54 in C6 cells (green) compared to an isotype control (blue).

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Normal biopsies of deparaffinized Mouse kidney tissue.

Immunohistochemistry: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Normal biopsies of deparaffinized Mouse lung tissue.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Normal biopsies of deparaffinized Mouse spleen tissue.

Flow Cytometry: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Staining of stimulated rat spleen cells with mouse anti-rat CD54 (Alexa488).

Flow Cytometry: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Analysis of CD54 in Ramos cells (green) compared to an isotype control (blue).

Flow Cytometry: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Analysis of CD54 in Raji cells (green) compared to an isotype control (blue).

Knockdown of RPN2 decreased luminal B BC cell dissemination in vivo. Luminal B ER+ T47D cells, transfected with negative control siRNA (siRNA-C) or siRNA targeting RPN2 (siRNA-RPN2) were injected + estradiol (E2) ± neutrophils (Neu) into zebrafish transgenic embryos with green fluorescent blood vessels & analyzed as described in materials & methods. (A) Migration in vitro (n = 6). (B)In vivo dissemination of transfected T47D in presence of E2 ± Neu (n = 23–26). Scale bar = 100 µm. (n = 23–26). (C) Western blot analysis for confirmation of siRNA-RPN2 transfection & VCAM-1, ICAM-1, & MUC-1 expression. (D) Focal adhesions area (n = 5–6). Scale bar = 10 µm. (E) Proliferation (n = 6). Representative images of zebrafish embryos with disseminated luminal B T47D BC cells & immunocytochemistry analysis of vinculin expression are shown. Arrows show disseminated T47D & arrowheads show focal adhesions. BV = blood vessels. Data are presented as mean ± SEM. Two-tailed Student’s t-test *P < 0.05, ***P < 0.001, ns, not significant. Data are represented of at least two independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33330095), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knockdown of U2AF1 decreased luminal A BC cell dissemination in vivo. Luminal A ER+ MCF-7 cells, transfected with negative control siRNA (siRNA-C) or siRNA targeting U2AF1 (siRNA-U2AF1) were injected in presence of estradiol (E2) ± neutrophils (Neu) into zebrafish transgenic embryos, with green fluorescent blood vessels, & analyzed as described in materials & methods. (A) Migration in vitro (n = 6–12). (B)In vivo dissemination in presence of E2 ± Neu (n = 38–41). Scale bar = 100 µm. (C) Western blot analysis for confirmation of siRNA-U2AF1 transfection & ICAM-1, VCAM-1, & MUC-1 expression. (D) Focal adhesion area (n = 7). Scale bar = 10 µm. (E) Proliferation in vitro (n = 12). Representative images of zebrafish embryos with disseminated MCF-7 cells & immunocytochemistry analysis of vinculin expression are shown. Arrows show disseminated MCF-7 cells & arrowheads show focal adhesions. BV = blood vessels. Data are presented as mean ± SEM. Two-tailed Student’s t-test *P < 0.05, **P < 0.01, ns, not significant. Data are represented of at least two independent experiments. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33330095), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Normalized gene & protein expression of ICAM-1. (A) qPCR results indicated no change in mRNA expression of ICAM-1 at 24 h post-overpressure but a significant increase at 48 h as compared to sham. (B) Subsequent Western blot results confirmed elevated ICAM-1 protein levels at 48 h compared to sham. *p < 0.05, Data are represented as mean ± SEM, gene: n = 7–9/group, protein: n = 8–9/group. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30853931), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Western Blot: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Fractionated, compared to single exposure, radiation more profoundly enhanced ICAM-1 expression. Representative (3 independent experiments) Western blot analysis & quantification of ICAM-1 levels in whole-cell lysates from nonirradiated (sham) & irradiated HUVECs 4 h & 24 h after exposure to (a) either five fractions of 2 Gy (5 × 2 Gy) or single exposure to 10 Gy & (b) either five fractions of 2.5 Gy (5 × 2.5 Gy) or single exposure to 12.5 Gy. Fractions delivered at 24-h intervals. beta -actin served as a loading control. c Ectopic expression of ICAM-1 after exposure to 0 Gy, five fractions of 2 Gy, or 10 Gy, as measured by flow cytometry (n = 3). d NF-kappa B activation after 4 h of exposure to 0 Gy, five fractions of 2 Gy, or 10 Gy (n = 2). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32382091), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

$Western Blot: ICAM-1/CD54 Antibody (1A29) [NBP2-22541] - Fractionated thoracic irradiation suppressed KLF2 & enhanced ICAM-1 levels in the lung. Quantification of KLF2 (a) & ICAM-1 (b) protein levels & representative Western blot analysis (c) in the lung tissue of mice (n = 6) at 24 h following 5 fractions of 4 Gy at 24 h intervals or single exposure to 20 Gy. beta -actin served as a loading control. KLF2 immunostaining in the lung tissue samples of sham irradiated (d), irradiated (e), & quantitation (f) at 24 h after exposure to 5 fractions of 4 Gy at 24 h intervals or single exposure to 20 Gy. ICAM-1 immunostaining in the lung tissue samples of sham irradiated (g), or irradiated (h), & quantitation (i) at 24 h after exposure to 5 fractions of 4 Gy at 24 h intervals or single exposure to 20 Gy. (n, number of independent experiments performed; a, significant statistical difference between nonirradiated & irradiated groups; b, significant statistical difference between fractionated irradiation & single exposure; *p < 0.05; **p < 0.01; ***p < 0.001). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32382091), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-22541AF350
Alexa Fluor 405	NBP2-22541AF405
Alexa Fluor 488	NBP2-22541AF488
Alexa Fluor 532	NBP2-22541AF532
Alexa Fluor 594	NBP2-22541AF594
Alexa Fluor 647	NBP2-22541AF647
Alexa Fluor 700	NBP2-22541AF700
Alexa Fluor 750	NBP2-22541AF750
Allophycocyanin	NBP2-22541APC
Allophycocyanin/Cy7	NBP2-22541APCCY7
Biotin	NBP2-22541B
CoraFluor 1	NBP2-22541CL1
DyLight 350	NBP2-22541UV
DyLight 405	NBP2-22541V
DyLight 488	NBP2-22541G
DyLight 550	NBP2-22541R
DyLight 594	NBP2-22541DL594
DyLight 650	NBP2-22541C
DyLight 680	NBP2-22541FR
DyLight 755	NBP2-22541IR
FITC	NBP2-22541F
HRP	NBP2-22541H
Janelia Fluor 525	NBP2-22541JF525
Janelia Fluor 549	NBP2-22541JF549
Janelia Fluor 585	NBP2-22541JF585
Janelia Fluor 635	NBP2-22541JF635
Janelia Fluor 646	NBP2-22541JF646
Janelia Fluor 669	NBP2-22541JF669
PE	NBP2-22541PE
PE/Atto594	NBP2-22541PEATT594
PE/Cy5.5	NBP2-22541PECY55
PE/Cy7	NBP2-22541PECY7
PerCP	NBP2-22541PCP
mFluor Violet 450 SE	NBP2-22541MFV450
mFluor Violet 500 SE	NBP2-22541MFV500
mFluor Violet 610 SE	NBP2-22541MFV610

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AF583

ICAM-1/CD54 Antibody [Unconjugated]

(26 Publications)

100 μg

AF583

Western blot shows lysates of rat liver tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Rat ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF583) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for ICAM-1/CD54 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ICAM‑1/CD54 was detected in perfusion fixed frozen sections of rat brain using Goat Anti-Rat ICAM‑1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF583) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Reactivity:

Species Glossary

Reactivity: Rt

Applications:

, ,

Applications: , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF583

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF583-SP

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BBA17

ICAM-1/CD54 Antibody [Unconjugated]

(18 Publications)

500 uL

BBA17

Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and Raji human Burkitt's lymphoma cell line. PVDF membrane was probed with 1:1000 µg/mL of Goat Anti-Human ICAM‑1/CD54 Polyclonal Antibody (Catalog # BBA17) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for ICAM‑1/CD54 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

ICAM-1/CD54 was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human ICAM-1/CD54 Polyclonal Antibody (Catalog # BBA17) at 1:600 dilution for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and human ICAM-1/CD54 Ramos human Burkitt's lymphoma cell line (KO). PVDF membrane was probed with 1:1000 µg/mL of Goat Anti-Human ICAM‑1/CD54 Polyclonal Antibody (Catalog # BBA17) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for ICAM‑1/CD54 at approximately 90 kDa (as indicated) in the parental Ramos human Burkitt's lymphoma cell line, but is not detectable in knockout Ramos human Burkitt's lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Requirement of NF-kappa B activation for cytokine-mediated reductions in TEER.A: Concentration-dependent inhibition of induction of ICAM-1 expression by I kappa K-beta -inhibitor Bay11, confirming the effect of Bay11 on TNF induction of NF-kappa B-dependent genes. Conditions are lane 1, DMSO Control; lanes, 2, 3, 4 and 5, Bay11 at 0.375, 0.75, 1.5 and 3.0 μM, respectively all after 0.8 ng/ml TNF for 16 hours. B: Effect of Bay11 concentration on the TNF-induced TEER decrease. ECIS analysis. X-axis: Duration of 0.8 ng/ml TNF treatment. Y-axis: TEER (ohms) normalized to basal barrier level prior to addition of TNF. TEER levels were not affected by these concentrations of Bay11 in the absence of TNF (not shown). The corrected basal TEER for this experiment was 69.9 ± 1.3 Ω·cm2. n = 6,6,6,6. C: Effects of SR-I kappa B dominant negative overexpression on HDMEC barrier responses. Upper panel: A time course of TNF treatment (1 ng/ml for 12 h) in control-transduced (black trace) and SR-I kappa B-transduced (red trace) HDMEC. n = 4,4. Lower panel: A time course of thrombin (1 U/ml for 12 h) in control-transduced (black trace) and SR-I kappa B-transduced (red trace) HDMEC. The corrected basal TEER for this experiment was 72.4 ± 0.9 Ω·cm2 for SR-I kappa B-transduced HDMEC and 80.0 ± 0.7 Ω·cm2 for vector control-transduced HDMEC. n = 3,3. Note that the phase 1 and phase 2 decreases initiated by TNF are markedly inhibited in SR-I kappa B-relative to control-transduced HDMEC but that thrombin-induced TEER decreases are similar in the same control- and SR-I kappa B-transduced HDMEC lines. D: Effects of Bay11 (used at 1 or 3 μM as labeled) on disruption of CL5 staining by 6 hours of TNF at 10 ng/ml. Morphometric measurements of TNF-induced disruption of CL5 junctional staining as described in the Methods (left). Immunofluorescence images representative of those used to assess the extent of disruption (right). Anti-CL5 (green), and anti-ICAM (red). Scale bar, 15 μm. E: Effects of SR-I kappa B transduction on disruption of CL5 staining by 6 hours of TNF at 10 ng/ml. Morphometric measurements (left) and representative immunofluorescence (right) images as in (D). Anti-CL5 (green), and anti-ICAM (red). Scale bar, 15 μm. Note that Bay 11 and SR-I kappa B each prevented the induction of ICAM-1 as well as the induction of a disrupted pattern of anti-CL5 immunofluorescence staining (arrows in D and E) by TNF. F: Effects of SR-I kappa B dominant negative overexpression on IL-1 beta -leak (ECIS). Note that IL-1 beta over a 12 hour time course was ineffective at decreasing TEER in SR-I kappa B-transduced HDMEC. The corrected basal TEER for this experiment was 64.5 ± 2.8 Ω·cm2 for SR-I kappa B-transduced HDMEC and 59.6 ± 1.1 Ω·cm2 for vector control-transduced HDMEC. n = 3,3. Representative of 3 (A, B and C) or 2 (D, E and F) independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120075), licensed under a CC-BY license. Not internally tested by R&D Systems.

Kinetics and dose response of distinct changes to HDMEC barriers induced by TNF and IL-beta .A: Relationship of the early TNF-induced TEER increase to basal TEER levels. A plot of the percent increase over basal TEER values (measured at the peak of the TNF-induced TEER increase, mean 0.7±0.01 hours; y-axis) vs. basal TEER (reported in ohms and read on a 96W20idf ECIS array; x-axis). TNF concentration was 20 ng/ml. The inverse correlation of basal TEER to the early TEER increase is statistically significant by a two-tailed Pearson analysis (p = 0.021 in 10 independent experiments). B: TNF induction of an early TEER rise and a bi-phasic TEER decrease. Labels indicate the peak of the early TEER increase and two distinct phases of TEER decrease (as nadirs to phases 1 and 2). X-axis, duration of incubation in 20 ng/ml TNF (units, hours); y-axis, in units of normalized TEER calculated as a ratio of TEER measurements taken post-TNF to the basal TEER level read before adding TNF, which is set at 1.0 (for a further explanation, please see Methods). The corrected basal TEER for this experiment was 63.7±1.2 Ω·cm2. n = 4,8 for vehicle, TNF. C: Relationship of TNF concentration to the decrease in TEER measured at the observed nadirs to phase 1 and phase 2. Example of data used to calculate EC50 values. Goodness of the non-linear regression curve fits are expressed as R-squared values. The corrected basal TEER for this experiment was 80.7± 0.8 Ω·cm2. D: Recovery of HDMEC barrier integrity relative to TNF concentration. TEER values show an inverse concentration-dependent recovery from phase 2 nadir levels (red trace) to pre-TNF basal levels in the continuous presence of TNF for 18 hours (black trace). The corrected basal TEER for this experiment was 57.5 ± 0.5 Ω·cm2. E: Effect of TNFR1 siRNA knockdown on TNF leak. Immunoblot analysis of siRNA silencing of TNFR1 expression confirmed by an inhibition of ICAM-1 expression (left) and ECIS analysis of the requirement for TNFR1 in TNF leak (right, y-axis TEER normalized to T0). The corrected basal TEER for this experiment was 75.6 ± 2.2 Ω·cm2 for TNFR1 siRNA-transfected HDMEC and 81.0 ± 1.2 Ω·cm2 for negative control siRNA-transfected HDMEC. Mean values are indicated by horizontal bars, n = 3,3) each at 10 hours of TNF at 0.8 ng/ml. MW, protein apparent molecular weight in kDa. F) Time course of discrete IL-beta -induced changes in TEER (ECIS plot). Note that like TNF, IL-beta (20 ng/ml) produced an initial small rise in TEER followed by two distinct phases of TEER decrease. The corrected basal TEER for this experiment was 72.0 ± 1.5 Ω·cm2. n = 3,3. Representative of 10 (A), 12 (B), 3 (C, D and F) or 2 (E) independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120075), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effects of TNF on the actin cytoskeleton and MLC phosphorylation.A) Analysis of the effects of TNF on actin/CL5 co-localization. Post-confluent HDMEC monolayers immunostained with anti-CL5 and phalloidin-stained for actin were imaged by fluorescence microscopy and analyzed for co-localization as described. Actin/CL5 co-localization was lost after TNF treatment for 8 hours at 0.8 ng/ml, resulting in a statistically significant difference in the Pearson correlation co-efficient (y-axis) by two-tailed t-test. B) Immunofluorescence microscopy representative of the data in Fig. 4. Co-localization of the cortical actin cytoskeleton with junctional CL5 in DMSO control HDMEC (arrow in left panel; phalloidin staining, red; anti-CL5, green) is dissociated by TNF (center panel), a change prevented by Bay11 (right panel). Scale bar, 15 μm. C) Time course of TNF-induced changes in MLC (Thr18/Ser19) phosphorylation and ICAM-1 levels measured by immunoblotting with controls for total MLC and for beta -actin. TNF treatment for the times indicated was at 10 ng/ml. D) Dose response of TNF-induced changes in phospho-MLC levels assessed by immunoblot analysis. HDMEC lysates were harvested at 6 hours of TNF. E) Effect of Bay11 on changes in P-MLC levels induced by TNF. Bay-11 was used at a 3 μM concentration. F) Effect of SR-I kappa B on changes in P-MLC levels induced by TNF. In E and F) TNF treatment was for 6 hours at 0.8 ng/ml. Note that Bay11 and SR-I kappa B each inhibit TNF-induced increases in ICAM-1 protein levels as well as MLC phosphorylation. Representative of 2 (B, E) or 3 (C, D, F) independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120075), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effects of ROCK inhibitors H-1152 and Y-27632 on TNF-induced MLC phosphorylation, actin and CL5 reorganization and the fall in TEER.A) Immunoblot analysis of the effects of H-1152 (top) or Y-27632 (bottom) concentration on induction of MLC phosphorylation at 6 hours of 0.8 ng/ml TNF. B) Fluorescence microscopy analysis of the effects of H-1152 or Y-27632 (at 10 μM concentrations) on TNF-induced actin re-organization. Actin visualized by phalloidin staining. Note that changes to the peripheral pattern of cortical actin at 6 hours of 0.8 ng/ml TNF treatment are inhibited by H-1152 and by Y-27632 (arrows). Scale bar, 15 μm. C) Top: Morphometric analysis of the effects of H-1152 (at concentrations of 2 or 20 μM, triangles and inverted triangles, respectively), and Y-27632 (at concentrations of 1 or 10 μM, squares and diamonds) on disruption of CL5 junctional staining at 6 hours of 0.8 ng/ml TNF. Open symbols, no TNF, closed symbols, plus TNF. Below: Examples of the morphometrically assessed immunofluorescence microscopy images. Note that CL5 staining disorganized by TNF in vehicle control appears condensed and contiguous in the presence of both ROCK inhibitors (arrows). Scale bar, 15 μm. D) Effects of H-1152 and Y-27632 on the phase 1 and phase 2 TEER decreases induced by TNF. Starting at T0, HDMEC plated on ECIS 96-well arrays received a 1 hour pre-treatment with vehicle (top panel, n = 4, 4) or with a ROCK inhibitor, either H-1152 (at 1 or 10 μM, middle panel, n = 6, 6) or Y-27632 (at 1 or 10 μM, bottom panel, n = 6, 6). One hour later 0.8 ng/ml TNF or (top panel only) vehicle was added, indicated by the vertical dashed lines. The corrected basal TEER for this experiment was 67.8 ± 0.4 Ω·cm2. E) Immunoblot analysis of the effects of H-1152 and Y-27632 on TNF-induced ICAM expression. Protein lysates were from HDMEC pre-treated with H-1152 (1 or 10 μM) or Y-27632 (1 or 10 μM) then TNF, 0.8 ng/ml for 6 h. Representative of 4 (A), 3 (B, D, E) and 2 (C) independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0120075), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

Host:

Goat Polyclonal

Host: Goat Polyclonal

Conjugate:

Unconjugated

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AF720

ICAM-1/CD54 Antibody [Unconjugated]

(1 Review)

(12 Publications)

100 μg

AF720

Western blot shows lysates of HUVEC human umbilical vein endothelial cells and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TNF-a (Catalog # 210-TA) for 24 hours. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF720) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ICAM-1/CD54 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ICAM-1/CD54 was detected in immersion fixed A431 human epithelial carcinoma cell line (left panel; positive stain) and A549 human lung carcinoma cell line (right panel; negative stain) using Sheep Anti-Human ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF720) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

Host:

Sheep Polyclonal

Host: Sheep Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Biotin	BAF720

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF720-SP

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NB500-318

ICAM-1/CD54 Antibody (MEM-111) - BSA Free

(7 Publications)

0.1 mg

NB500-318

Western Blot: ICAM-1/CD54 Antibody (MEM-111) [NB500-318] - Of CD54 expression in activated (A) and nonactivated (B) Huvec cells by antibody MEM-111. Lower bands represent tubulin as a loading control.

Flow Cytometry: ICAM-1/CD54 Antibody (MEM-111) [NB500-318] - Analysis using the FITC conjugate of NB500-318. Staining of Human peripheral blood lymphocytes with MEM-111 FITC.

Flow Cytometry: ICAM-1/CD54 Antibody (MEM-111) [NB500-318] - Analysis using PE conjugate of NB500-318. A cell surface stain was performed on THP-1 cells with ICAM-1/CD54 antibody (MEM-111) NB500-318 (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 1:100 for 20 minutes at room temperature. Both antibodies were conjugated to PE.

Reactivity:

Hu, Rt, Bv

Species Glossary

Reactivity: Hu, Rt, Bv

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB500-318AF350
Alexa Fluor 405	NB500-318AF405
Alexa Fluor 488	NB500-318AF488
Alexa Fluor 532	NB500-318AF532
Alexa Fluor 594	NB500-318AF594
Alexa Fluor 647	NB500-318AF647
Alexa Fluor 700	NB500-318AF700
Alexa Fluor 750	NB500-318AF750
Allophycocyanin	NB500-318APC
Allophycocyanin/Cy7	NB500-318APCCY7
Biotin	NB500-318B
CoraFluor 1	NB500-318CL1
DyLight 350	NB500-318UV
DyLight 405	NB500-318V
DyLight 488	NB500-318G
DyLight 550	NB500-318R
DyLight 594	NB500-318DL594
DyLight 650	NB500-318C
DyLight 680	NB500-318FR
DyLight 755	NB500-318IR
FITC	NB500-318F
HRP	NB500-318H
Janelia Fluor 525	NB500-318JF525
Janelia Fluor 549	NB500-318JF549
Janelia Fluor 585	NB500-318JF585
Janelia Fluor 635	NB500-318JF635
Janelia Fluor 646	NB500-318JF646
Janelia Fluor 669	NB500-318JF669
PE	NB500-318PE
PE/Atto594	NB500-318PEATT594
PE/Cy5.5	NB500-318PECY55
PE/Cy7	NB500-318PECY7
PerCP	NB500-318PCP
mFluor Violet 450 SE	NB500-318MFV450
mFluor Violet 500 SE	NB500-318MFV500
mFluor Violet 610 SE	NB500-318MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB500-318-25ug

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MAB720

ICAM-1/CD54 Antibody (14C11) [Unconjugated]

(5 Publications)

100 μg

MAB720

ICAM-1/CD54 was detected in immersion fixed A431 human epithelial carcinoma cell line using Mouse Anti-Human ICAM-1/CD54 Monoclonal Antibody (Catalog # MAB720) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

, ,

Applications: , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB720-SP
Carrier Free	MAB720-500

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MAB796

ICAM-1/CD54 Antibody (166623)

(6 Publications)

100 μg

Reactivity:

Species Glossary

Reactivity: Mu

Applications:

Applications: ,

Host:

Rat Monoclonal

Host: Rat Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Phycoerythrin	FAB796P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	FAB796P
Carrier Free	MAB796-SP
Carrier Free	MAB796-500

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NBP2-22540

ICAM-1/CD54 Antibody (3E2B) - Azide and BSA Free

(1 Publication)

500μg

NBP2-22540

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody (3E2B) [NBP2-22540] - Normal biopsies of deparaffinized Mouse spleen tissue.

Flow Cytometry: ICAM-1/CD54 Antibody (3E2B) [NBP2-22540] - Analysis of CD54 in mouse splenocytes (green) compared to an isotype control (blue).

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody (3E2B) [NBP2-22540] - Normal biopsies of deparaffinized Mouse kidney tissue.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody (3E2B) [NBP2-22540] - Normal biopsies of deparaffinized Mouse lung tissue.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , , , , , ,

Applications: , , , , , , ,

Host:

Hamster Monoclonal

Host: Hamster Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-22540AF350
Alexa Fluor 532	NBP2-22540AF532
Alexa Fluor 594	NBP2-22540AF594
Alexa Fluor 750	NBP2-22540AF750
DyLight 594	NBP2-22540DL594
Janelia Fluor 549	NBP2-22540JF549
Janelia Fluor 646	NBP2-22540JF646
mFluor Violet 450 SE	NBP2-22540MFV450
mFluor Violet 500 SE	NBP2-22540MFV500
mFluor Violet 610 SE	NBP2-22540MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
Azide and BSA Free	NBP2-22540-100ug

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NBP1-88700

ICAM-1/CD54 Antibody

(2 Publications)

0.1 ml

NBP1-88700

Western Blot: ICAM-1/CD54 Antibody [NBP1-88700] - Analysis in human lung tissue.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody [NBP1-88700] - Staining of human kidney shows moderate membranous positivity in cells in glomeruli.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody [NBP1-88700] - Staining of human lymph node shows moderate membranous positivity.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody [NBP1-88700] - Staining of human lung shows moderate membranous positivity in pneumocytes.

Immunohistochemistry-Paraffin: ICAM-1/CD54 Antibody [NBP1-88700] - Staining of human skeletal muscle shows no positivity in myocytes as expected.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

Applications: ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Validated by:

Validated by:

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NB110-58732

ICAM-1/CD54 Antibody (1H4) - Azide and BSA Free

0.1 mg

NB110-58732

Flow Cytometry: ICAM-1/CD54 Antibody (1H4) - Azide Free [NB110-58732] - Fig. 1. Surface staining of U937 human histiocytic lymphoma cell line with anti-human CD54 (1H4) PE. Total viable cells were used for analysis. Image using the Biotin form of this antibody.

Flow Cytometry: ICAM-1/CD54 Antibody (1H4) - Azide Free [NB110-58732] - Staining of human peripheral blood with anti-CD54 (1H4) APC.

Flow Cytometry: ICAM-1/CD54 Antibody (1H4) - Azide Free [NB110-58732] - Staining of human peripheral blood with anti-CD54 (1H4) Alexa Fluor (R) 700.

Flow Cytometry: ICAM-1/CD54 Antibody (1H4) - Azide Free [NB110-58732] - Surface staining of human peripheral blood with anti-CD54 (1H4) PerCP. Image from the PE version of this antibody.

Reactivity:

Species Glossary

Reactivity: Hu

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB110-58732AF350
Alexa Fluor 405	NB110-58732AF405
Alexa Fluor 488	NB110-58732AF488
Alexa Fluor 532	NB110-58732AF532
Alexa Fluor 594	NB110-58732AF594
Alexa Fluor 647	NB110-58732AF647
Alexa Fluor 700	NB110-58732AF700
Alexa Fluor 750	NB110-58732AF750
Allophycocyanin	NB110-81732
Allophycocyanin/Cy7	NB110-58732APCCY7
Biotin	NB500-584
CoraFluor 1	NB110-58732CL1
DyLight 350	NB110-58732UV
DyLight 405	NB110-58732V
DyLight 488	NB110-58732G
DyLight 550	NB110-58732R
DyLight 594	NB110-58732DL594
DyLight 650	NB110-58732C
DyLight 680	NB110-58732FR
DyLight 755	NB110-58732IR
FITC	NB500-583-100Tests
HRP	NB110-58732H
Janelia Fluor 525	NB110-58732JF525
Janelia Fluor 549	NB110-58732JF549
Janelia Fluor 585	NB110-58732JF585
Janelia Fluor 635	NB110-58732JF635
Janelia Fluor 646	NB110-58732JF646
Janelia Fluor 669	NB110-58732JF669
PE	NB110-58732PE
PE	NB500-582-100Tests
PE/Atto594	NB110-58732PEATT594
PE/Cy5.5	NB110-58732PECY55
PE/Cy7	NB110-58732PECY7
PerCP	NB110-58732PCP
mFluor Violet 450 SE	NB110-58732MFV450
mFluor Violet 500 SE	NB110-58732MFV500
mFluor Violet 610 SE	NB110-58732MFV610

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

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ICAM-1/CD54 Antibodies

ICAM-1/CD54 Antibody [Unconjugated]

ICAM-1/CD54 Antibody (BBIG-I1 (11C81)) [Unconjugated]

ICAM-1/CD54 Antibody (1A29)

ICAM-1/CD54 Antibody [Unconjugated]

ICAM-1/CD54 Antibody [Unconjugated]

ICAM-1/CD54 Antibody [Unconjugated]

ICAM-1/CD54 Antibody (MEM-111) - BSA Free

ICAM-1/CD54 Antibody (14C11) [Unconjugated]

ICAM-1/CD54 Antibody (166623)

ICAM-1/CD54 Antibody (3E2B) - Azide and BSA Free

ICAM-1/CD54 Antibody

ICAM-1/CD54 Antibody (1H4) - Azide and BSA Free