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AF2085

Brachyury Antibody [Unconjugated]

(2 Reviews)

(262 Publications)

100 μg

AF2085

Western blot shows 10 ng of Recombinant Human Brachyury (amino acids Ser2-Glu202) and Recombinant Human TBX6. PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Brachyury at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Western blot shows lysates of NCI‑H460 human large cell lung carcinoma cell line and MCF 10A human breast epithelial cell line (negative control). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Brachyury at approximately 50 kDa (as indicated). GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Brachyury was detected in immersion fixed differentiated human embryonic stem cells using 10 µg/mL Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) for 3 hours at room temperature. Cells were stained (green) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Brachyury was detected in immersion fixed BG01V human embryonic stem cells differentiated into mesoderm using Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counter-stained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Brachyury was detected in immersion fixed frozen sections of embryonic mouse notochord (E9.5) using 10 µg/mL Goat Anti-Human Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Mesoderm-differentiated BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Brachyury/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The VEGF promoter was detected by standard PCR.

NCI-H460 cells were stained with Goat Anti-Human/Mouse Brachyury Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2085, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Generation and characterization of systemic lupus erythematosus (SLE)‐specific human‐induced pluripotent stem cells (hiPSCs). (A) Dermal fibroblasts derived from patient with SLE were reprogrammed into iPSCs using Sendai virus vectors and three clones (#1, #2 and #3) were characterized. RT‐PCR confirms the loss of transgenes in hiPSCs‐SLE (lanes 1, 2 and 3), presence (lane 4) in infected fibroblasts (ipF‐SLE) and absence of Sendai viral transgenes in parental fibroblasts (pF‐SLE) (lane 5). Full‐length gels are presented in File S2. (B) The hiPSCs‐SLE colonies expressed alkaline phosphatase. Scale bar, 500 μm. (C) RT‐qPCR analysis of pluripotency genes OCT4, NANOG, SOX2 and REX1 was performed in fibroblasts and in hiPSCs derived from patient with SLE. All expression values are normalized to GAPDH and relative donor fibroblasts. Data are mean ± SEM and all statistical analysis was made between hiPSCs‐SLE clones and relative fibroblasts by Student's t test showing P‐values ≤ .05 in each comparison. (D) PluriTest assays combines novelty score (blue) on x‐axis and pluripotency score (red) on y‐axis. hiPSCs‐SLE localize in the red cloud suggesting the empirical distribution of pluripotent cells compared to non‐pluripotent blue cloud. (E) Representative images of M‐FISH staining show normal karyotypes of hiPSCs‐SLE clones. (F) Immunofluorescence analysis of pluripotent stem cell markers Nanog (green), Oct4 (red) and co‐staining with DAPI (blue) in hiPSCs‐SLE. Scale bar, 50 μm. (G) Representative images of floating and adherent EBs derived from hiPSCs‐SLE at differentiation day 8 and 18, respectively. Scale bar, 500 μm. (H) RT‐qPCR results confirm the capability of hiPSCs‐SLE to differentiate into all three germ layers. The expression levels of GATA4, HAND1 and PAX6 in EBs are relative to undifferentiated hiPSCs. All expression values are normalized to GAPDH and relative hiPSCs. Data are mean ± SEM and all statistical analysis was made between EBs‐SLE and relative hiPSCs‐SLE clones by Student's t test showing P‐values ≤ .05 in each comparison. (I) Immunostaining of Otx2 (ectoderm marker), Sox17 (endoderm marker), Brachyury (mesoderm marker) and co‐staining with DAPI (blue) in EBs derived from hiPSCs‐SLE. Scale bar, 50 μm Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31536674), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reactivity:

Hu, Mu

Species Glossary

Reactivity: Hu, 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
Alexa Fluor 488	IC2085G
Allophycocyanin	IC2085A
Biotin	BAF2085
NL557	NL2085R
Phycoerythrin	IC2085P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	IC2085P
Carrier Free	IC2085A
Carrier Free	IC2085G
Carrier Free	NL2085R
Carrier Free	AF2085-SP

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MAB20851

Brachyury Antibody (1161B) [Unconjugated]

(10 Publications)

100 μg

MAB20851

Brachyury was detected in immersion fixed BG01V human embryonic stem cells differentiated into mesoderm using Rabbit Anti-Human Brachyury Monoclonal Antibody (Catalog # MAB20851) at a 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	MAB20851-SP

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NL2085R

Brachyury Antibody [NL557]

0.5 mL

NL2085R

Brachyury was detected in E9.5 mouse embryo sections using Human Brachyury NL557 conjugated Antibody (Catalog# NL2085R). Embryos were fixed in 4% paraformaldehyde, and blocked with PBS containing 10% normal donkey serum, 0.1% Triton® X 100, and 1% BSA. After blocking, sections were incubated with NL557 conjugated antibody at a final concentration of 1X (1:10 dilution) in blocking buffer for 3 hours at room temperature in the dark. Between each step, cells were washed with PBS containing BSA.

Reactivity:

Hu

Species Glossary

Reactivity: Hu

Applications:

Applications:

Host:

Goat Polyclonal

Host: Goat Polyclonal

Conjugate:

NL557

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 488	IC2085G
Allophycocyanin	IC2085A
Biotin	BAF2085
Phycoerythrin	IC2085P

Available Formulations

Formulation	Catalog #	Availability	Price
Carrier Free	AF2085
Carrier Free	IC2085P
Carrier Free	IC2085A
Carrier Free	IC2085G
Carrier Free	AF2085-SP

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