Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [5]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- AF5078 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Sheep Polyclonal RelA/NFkB p65 Antibody
- Antibody type
- Polyclonal
- Description
- Antigen Affinity-purified. Detects human, mouse, and rat RelA/NF kappa B p65 in Western blots.
- Reactivity
- Human, Mouse, Rat
- Host
- Sheep
- Conjugate
- Unconjugated
- Isotype
- IgG
- Vial size
- 100 ug
- Concentration
- LYOPH
- Storage
- Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
Submitted references Characterization of short range DNA looping in endotoxin-mediated transcription of the murine inducible nitric-oxide synthase (iNOS) gene.
Guo H, Mi Z, Kuo PC
The Journal of biological chemistry 2008 Sep 12;283(37):25209-17
The Journal of biological chemistry 2008 Sep 12;283(37):25209-17
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Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of Human RelA/NF kappa B p65 by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, Daudi human Burkitt's lymphoma cell line, and LNCaP human prostate cancer cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for RelA/NF kappa B p65 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of Human and Mouse RelA/NF kappa B p65 by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, HeLa human cervical epithelial carcinoma cell line, and Neuro-2A mouse neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for RelA/NF kappa B p65 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of Human RelA/NF kappa B p65 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for RelA/NF kappa B p65 at approximately 68 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of Human, Mouse, and Rat RelA/NF?B p65 by Simple WesternTM. Detection of Human, Mouse, and Rat RelA/NF?B p65 by Simple WesternTM.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of Mouse and Rat RelA/NF?B p65 by Western Blot. Detection of Mouse and Rat RelA/NF?B p65 by Western Blot.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- RelA/NF kappa B p65 in Human Squamous Cell Carcinoma. RelA/NF kappa B p65 was detected in immersion fixed paraffin-embedded sections of human squamous cell carcinoma using Sheep Anti-Human/Mouse RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Detection of RelA/NF kappa B p65-regulated Genes by Chromatin Immuno-precipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for overnight was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. RelA/NF kappa B p65/DNA complexes were immunoprecipitated using 5 μg Sheep Anti-Human RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The p21 promoter was detected by standard PCR.