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AF2720

Tie-2 [p Tyr992] Antibody [Unconjugated]

(1 Review)

(32 Publications)

50 μg

AF2720

Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line transfected with mouse Tie-2 and untreated (-) or treated (+) with 600 ng/mL Recombinant Human Angiopoietin‑1 (Catalog # 923-AN) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Phospho-Tie‑2 (Y992) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2720) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Tie‑2 (Y992) at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Tie‑2 was detected in immersion fixed paraffin-embedded sections of human kidney tissue using Rabbit Anti-Human/Mouse Phospho-Tie‑2 (Y992) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2720) at 1 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC003) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

HUVEC human umbilical vein endothelial cells were unstimulated (light orange filled histogram) or treated with 100 μM pervanadate for 15 minutes (dark orange filled histogram), then stained with Human/Mouse Phospho-Tie‑2 (Y992) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF2720) or control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Pericyte-derived Angpt1 controls alveologenesis. a RT-qPCR analysis of Angpt1 and Tie2/Tek expression in freshly sorted lung GFP+, CD31+ or EpCAM+ cells from P7 Pdgfrb(BAC)-CreERT2 R26-mT/mG mice. Data represents mean ± s.e.m. (n = 4 mice). b High magnification images of P10 Angpt1GFP lungs stained for GFP (green), PDGFR beta (red), and PDGFR alpha (blue). Arrows indicate GFP and PDGFR beta double positive pericytes. Scale bar, 15 µm. c RT-qPCR analysis of Angpt1 expression in freshly sorted PDGFR beta + cells from P7 Yap1,Wwtr1iPCKO and control lungs. Data represents mean ± s.e.m. (n = 4 mice, two-tailed unpaired t-test). dAngpt1 expression in cultured Verteporfin (VP)-treated (48 h) and control pericytes. Data represents mean ± s.e.m. (n = 4, Welch’s t-test). e Expression of the indicated transcripts in freshly sorted CD31+ cells from P7 Yap1,Wwtr1iPCKO and control lungs. Data represents mean ± s.e.m. (n = 4 mice, NS not significant, two-tailed unpaired t-test). f–h Western blot analysis of Angpt1 protein (f; n = 2 controls and 4 mutant mice) and of total and phospho-Tie2 (pTie2) in P12 Yap1,Wwtr1iPCKO and control total lung lysates (g, n = 3 controls and 5 mutants). Molecular weight marker (kDa) is indicated. Relative quantification of signals is shown in h. Two-tailed unpaired t-test. i Scheme showing the time points of tamoxifen administration and analysis for Angpt1iPCKO mice. j, k 3D reconstruction confocal images of P12 Angpt1iPCKO and littermate control lungs stained for AQP5 (green), PDGFR beta (red), and PECAM1 (blue). Panels in k show higher magnification of PECAM1 staining. Scale bar, 50 µm (j) and 30 µm (k). l Quantitation of airspace volume in P12 Angpt1iPCKO and littermate control lung sections with 3D reconstruction surface images. Data represents mean ± s.e.m. (n = 4 mice; p < 0.0001, two-tailed unpaired t-test). m 3D reconstruction confocal images of P12 Angpt1iPCKO and littermate control lungs stained for alpha SMA (red) and tropoelastin (blue). Scale bar, 50 µm. n Quantitation of staining intensity for alpha SMA or tropoelastin shown in m. Intensity was normalized to the size of the parenchymal region. Data represents mean ± s.e.m. (n = 4 mice; NS not significant, two-tailed unpaired t-test). o Schematic summary of findings. Pulmonary pericytes regulate ECs and alveolar epithelial cells via angiocrine factors such as angiopoietin-1 and HGF Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29934496), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, Mu

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2720-SP

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NBP1-44894

EGFR [p Tyr992] Antibody (EM-12) - BSA Free

0.1 mg

NBP1-44894

Western Blot: EGF R/ErbB1 [p Tyr992] Antibody (EM-12) [NBP1-44894] - Analysis of EGFR (phospho-Tyr992) by mouse monoclonal antibody EM-12 in EGF-treated A431 (A), CALU-3 (B), MCF-7 (C), Jurkat (D) and Ramos (E) cell lines (reduced conditions).

Immunoprecipitation: EGF R/ErbB1 [p Tyr992] Antibody (EM-12) [NBP1-44894] - EGFR from EGF-treated A431 cells by monoclonal antibodies EM-12 (A), EM-13 (B), a commercial anti-EGFR polyclonal antibody (C) and anti-EGFR monoclonal (D). The precipitates were immunoblotted with EM-12 or EM-13 antibody, and goat anti-mouse-HRP.

Immunoprecipitation: EGF R/ErbB1 [p Tyr992] Antibody (EM-12) [NBP1-44894] - EGFR from EGF-treated A431 cells by phosphospecific monoclonal antibodies EM-12 (A), EM-13 (B), a commercial anti-EGFR polyclonal antibody (C) and anti-EGFR monoclonal (D). The precipitates were immunoblotted with a commercial anti-EGFR polyclonal and goat anti-rabbit-HRP.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

, , ,

Applications: , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-44894AF350
Alexa Fluor 405	NBP1-44894AF405
Alexa Fluor 488	NBP1-44894AF488
Alexa Fluor 532	NBP1-44894AF532
Alexa Fluor 594	NBP1-44894AF594
Alexa Fluor 647	NBP1-44894AF647
Alexa Fluor 700	NBP1-44894AF700
Alexa Fluor 750	NBP1-44894AF750
Allophycocyanin	NBP1-44894APC
Allophycocyanin/Cy7	NBP1-44894APCCY7
Biotin	NBP1-44894B
DyLight 350	NBP1-44894UV
DyLight 405	NBP1-44894V
DyLight 488	NBP1-44894G
DyLight 550	NBP1-44894R
DyLight 594	NBP1-44894DL594
DyLight 650	NBP1-44894C
DyLight 680	NBP1-44894FR
DyLight 755	NBP1-44894IR
FITC	NBP1-44894F
HRP	NBP1-44894H
Janelia Fluor 549	NBP1-44894JF549
Janelia Fluor 646	NBP1-44894JF646
PE	NBP1-44894PE
PE/Atto594	NBP1-44894PEATT594
PE/Cy5.5	NBP1-44894PECY55
PE/Cy7	NBP1-44894PECY7
PerCP	NBP1-44894PCP
mFluor Violet 450 SE	NBP1-44894MFV450
mFluor Violet 500 SE	NBP1-44894MFV500
mFluor Violet 610 SE	NBP1-44894MFV610

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NBP3-13265

EGFR [p Tyr992] Antibody

100 ul

NBP3-13265

Western Blot: EGFR [p Tyr992] Antibody [NBP3-13265] - Untreated (-) and treated (+) A431 whole cell extract (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with EGFR (phospho Tyr992) antibody (NBP3-13265) diluted at 1:2000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: EGFR [p Tyr992] Antibody [NBP3-13265] - EGFR (phospho Tyr992) antibody detects EGFR (phospho Tyr992) protein at cell membrane by immunofluorescent analysis. Sample: A431 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: EGFR (phospho Tyr992) protein stained by EGFR (phospho Tyr992) antibody (NBP3-13265) diluted at 1:500. Blue: Hoechst 33342 staining.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

,

Applications: ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NBP3-13228

EGFR [p Tyr992] Antibody

100 ul

NBP3-13228

Western Blot: EGFR [p Tyr992] Antibody [NBP3-13228] - Untreated (-) and treated (+) A431 whole cell extract (30 ug) were separated by 7.5% SDS-PAGE, and the membrane was blotted with EGFR (phospho Tyr992) antibody (NBP3-13228) diluted at 1:2000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: EGFR [p Tyr992] Antibody [NBP3-13228] - EGFR (phospho Tyr992) antibody detects EGFR (phospho Tyr992) protein at cell membrane by immunofluorescent analysis. Sample: A431 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: EGFR (phospho Tyr992) protein stained by EGFR (phospho Tyr992) antibody (NBP3-13228) diluted at 1:500. Blue: Hoechst 33342 staining.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

,

Applications: ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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