Human/Mouse/Rat RelA/NF kappa B p65 Antibody

Catalog # Availability Size / Price Qty
AF5078
AF5078-SP
Detection of Human RelA/NF kappa B p65 by Western Blot.
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Product Details
Citations (3)
FAQs
Supplemental Products
Reviews (2)

Human/Mouse/Rat RelA/NF kappa B p65 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat RelA/NF kappa B p65 in Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human RelA/NF kappa B p65 isoform 1
Asn456-Ser551
Accession # Q04206
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
20 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 106 cells
See below
Immunocytochemistry
5-15 µg/mL
See below
Knockout Validated
RelA/NF kappa B p65 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in RelA/NF kappa B p65 knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection of Human RelA/NF?B p65 antibody by Western Blot. View Larger

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/Mouse/Rat 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.

Western Blot Detection of Human and Mouse RelA/NF?B p65 antibody by Western Blot. View Larger

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/Mouse/Rat 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.

Western Blot Detection of Mouse and Rat RelA/NF?B p65 antibody by Western Blot. View Larger

Detection of Mouse and Rat RelA/NF kappa B p65 by Western Blot. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line and C6 rat glioma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse 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 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Chromatin Immunoprecipitation (ChIP) Detection of RelA/NF kappa B p65-regulated Genes by Chromatin Immuno-precipitation. View Larger

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/Mouse/Rat 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 # [[catalogNumber:BAF016)]]. Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # [[catalogNumber:MAG999]]) and DNA was purified using chelating resin solution. Thep21promoter was detected by standard PCR.

Immunocytochemistry RelA/NF?B p65 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

RelA/NF kappa B p65 in HeLa Human Cell Line. RelA/NF kappa B p65 was detected in immersion fixed HeLa human cervical epithelial carcinoma cells untreated (upper panel) or treated (lower panel) with 20 ng/mL Recombinant Human TNF-alpha (Catalog # 210-TA) for 10 minutes using Sheep Anti-Human/Mouse/Rat RelA/NF kappa B p65 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5078) at 10 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 cytoplasm in untreated cells and nuclei in treated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry RelA/NF?B p65 antibody in Human Squamous Cell Carcinoma by Immunohistochemistry (IHC-P). View Larger

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/Rat 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.

Simple Western Detection of Human, Mouse, and Rat RelA/NF?B p65 antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human, Mouse, and Rat RelA/NF kappa B p65 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, Neuro-2A mouse neuroblastoma cell line, and C6 rat glioma cell line, loaded at 0.5 mg/mL. A specific band was detected for RelA/NF kappa B p65 at approximately 65 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human/Mouse/Rat 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.

Knockout Validated Western Blot Shows Human RelA/NF?B p65 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human RelA/NF kappa B p65 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and RelA/NF kappa B p65 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat 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 65 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: RelA/NFkB p65

RelA belongs to a family of transcription factors (NF kappa B (nuclear factor kappa from B cells) complex) that play a fundamental role in inflammatory and immune responses. The NF kappa B complex is composed of a heterodimer of a Rel family member (RelA, c-Rel, RelB) and either NF kappa B1 or NF kappa B2 subunits. RelA and NF kappa B1 are the most common heterodimeric pair. The NF kappa B complex is sequestered in the cytoplasm by inhibitory I kappa B proteins. Upon cellular activation, the ubiquitin-proteosome pathway degrades the I kappa B proteins allowing the NF kappa B complex to translocate to the nucleus and activate gene transcription.

Long Name
v-rel Reticuloendotheliosis Viral Oncogene Homolog A
Entrez Gene IDs
5970 (Human)
Alternate Names
MGC131774; nf kb p65; NFkB p65; NF-kB p65; NFKB3; NFKB3v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor ofkappa light polypeptide gene enhancer in B-cells 3 (p65)); Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3transcription factor p65; p65; p65RelA; rela p65; RelA; v-rel reticuloendotheliosis viral oncogene homolog A (avian); v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappalight polypeptide gene enhancer in B-cells 3, p65

Product Datasheets

Citations for Human/Mouse/Rat RelA/NF kappa B p65 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Anagliptin prevents lipopolysaccharide (LPS)- induced inflammation and activation of macrophages
    Authors: F Yu, W Tian, J Dong
    International immunopharmacology, 2022;104(0):108514.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. TET repression and increased DNMT activity synergistically induce aberrant DNA methylation
    Authors: H Takeshima, T Niwa, S Yamashita, T Takamura-E, N Iida, M Wakabayash, S Nanjo, M Abe, T Sugiyama, YJ Kim, T Ushijima
    J. Clin. Invest., 2020;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  3. Characterization of short range DNA looping in endotoxin-mediated transcription of the murine inducible nitric-oxide synthase (iNOS) gene.
    Authors: Guo H, Mi Z, Kuo PC
    J. Biol. Chem., 2008;283(37):25209-17.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: ChIP

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Human/Mouse/Rat RelA/NF kappa B p65 Antibody
By Anonymous on 10/24/2019
Application: ELISA Sample Tested: xenograft PD sample Species: Human

Human/Mouse RelA/NF kappa B p65 Antibody
By Anonymous on 08/23/2018
Application: Western Blot Sample Tested: Human fibroblast Species: Human