Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 532301
View all RelA/NFkB p65 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human RelA/NF kappa B p65 isoform 1
Asn456-Ser551
Accession # Q04206

Specificity

Detects human, mouse, and rat RelA/NF kappa B p65 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human/Mouse/Rat RelA/NF kappa B p65 Antibody

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

Detection of Human RelA/NF kappa B p65 by Western Blot.

Western Blot shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 2 µg/ml of Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 Western Blot Buffer Group 1.

Detection of Mouse RelA/NF kappa B p65 by Western Blot.

Western Blot shows lysates of BaF3 mouse pro-B cell line. PVDF membrane was probed with 2 µg/ml of Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 Western Blot Buffer Group 1.
Detection of Rat RelA/NF?B p65 antibody by Western Blot.

Detection of Rat RelA/NF kappa B p65 by Western Blot.

Western blot shows lysates of C6 rat glioma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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.
RelA/NF?B p65 antibody in K562 Human Cell Line by Immunocytochemistry (ICC).

RelA/NF kappa B p65 in K562 Human Cell Line.

RelA/NF kappa B p65 was detected in immersion fixed K562 human chronic myelogenous leukemia cell line using Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078) at 10 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). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of RelA antibody in HeLa Human Cell Line antibody by Flow Cytometry.

Detection of RelA in HeLa Human Cell Line by Flow Cytometry.

HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformadehyde and permeabilized with methanol.
Western Blot Shows Human RelA/NF?B p65 Antibody Specificity by Using Knockout Cell Line.

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 2 µg/mL of Mouse Anti-Human/Mouse/Rat RelA/NF kappa B p65 Monoclonal Antibody (Catalog # MAB5078) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). 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.
Detection of Human RelA/NFkB p65 by Western Blot

Detection of Human RelA/NFkB p65 by Western Blot

Non-classical and intermediate monocytes express high levels of NF-kappa B (p65) and membrane-bound IL-1 alpha.a Western blot analysis of total p65 and GAPDH protein levels in the three monocyte subsets. b Quantification of Western blot data shown in (a): p65 protein level was normalized to GAPDH (loading control) and expressed as a fold change with respect to CL subset. The data represent the means ± SD; n = 3. c Relative levels of phosphorylated-p65 (p-p65), measured by flow cytometry. Each line represents one donor; n = 7. d IL-1 alpha secretion by the three monocyte subsets, measured by Luminex assay. The data represent the means ± SD; n = 3. e–f Relative expression levels of membrane-bound IL-1 alpha (e) and cytoplasmic IL-1 alpha (f), analyzed by flow cytometry. Each line represents one donor; n = 9. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. CL: classical, ITM: intermediate, NC: non-classical, MFI: median fluorescence intensity Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/s41419-018-0327-1), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human RelA/NFkB p65 by Western Blot

Detection of Human RelA/NFkB p65 by Western Blot

Tumor‐derived EVs activate the NF kappa B pathway. Primary CD14+ monocytes were incubated with TEVs isolated from conditioned PCI‐1 supernatants. (A) Lysates were generated at different timepoints of incubation and tested for I kappa B and phosphorylated P‐I kappa B. Tubulin was used as a loading control. (B) Primary monocytes were incubated with complete PCI‐1 supernatant (SN), in TEV‐depleted SN (depl.) or with isolated TEV for 120 min, nuclei were isolated and the translocation of NF kappa B‐p65 was tested with an immunoblot. Actin was used as a loading control, and the cytoplasmic fraction (p65/cyto) was used to confirm the specificity of the translocation. (C) Monocytes were incubated as in (B), and the binding capacity of NF kappa B p50 to the DNA was quantified with a NF kappa B binding assay kit. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29601673), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RelA/NF kappa B p65 by Western Blot

Detection of RelA/NF kappa B p65 by Western Blot

Impact of resveratrol or/and ASO against beta 1-integrin on TME-promoted activation of metastasis and apoptosis parameters in CRC cells. HCT116 (A) and RKO (B) derived from 3D-alginate cultures were grown untreated or treated with 5 µM resveratrol alone or in combination with 0.5 µM beta 1-SO or 0.5 µM beta 1-ASO (x-axis) and probed with antibodies against CXCR4, FAK, p-FAK, NF-kappa B, p65-NF-kappa B and cleaved caspase-3. Loading control: beta -actin. Densitometric units complementing Western blot results (y-axis). For densitometric analysis, data were compared to TME control: *p < 0.05 and **p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36120305), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of RelA/NF kappa B p65 by Western Blot

Detection of RelA/NF kappa B p65 by Western Blot

Impact of resveratrol or/and ASO against beta 1-integrin on TME-promoted activation of metastasis and apoptosis parameters in CRC cells. HCT116 (A) and RKO (B) derived from 3D-alginate cultures were grown untreated or treated with 5 µM resveratrol alone or in combination with 0.5 µM beta 1-SO or 0.5 µM beta 1-ASO (x-axis) and probed with antibodies against CXCR4, FAK, p-FAK, NF-kappa B, p65-NF-kappa B and cleaved caspase-3. Loading control: beta -actin. Densitometric units complementing Western blot results (y-axis). For densitometric analysis, data were compared to TME control: *p < 0.05 and **p < 0.01. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36120305), licensed under a CC-BY license. Not internally tested by R&D Systems.

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

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed K562 human chronic myelogenous leukemia cell line

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line, fixed with paraformaldehyde and permeabilized with methanol

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.

Western Blot

2 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line, HeLa human cervical epithelial carcinoma cell line, BaF3 mouse pro-B cell line, and C6 rat glioma cell line

Reviewed Applications

Read 2 reviews rated 5 using MAB5078 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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Background: RelA/NFkB p65

RelA p65 (v-rel reticuloendotheliosis viral oncogene homolog A) is a 65 kDa member of the NF kappa B family of nuclear transcription factors. Dimers of p65 with the p50 subunit are the most common form of the NF kappa B transcription factor, but dimers with it or other family members can also occur. Upon activation, RelA p65 forms an heterotetramer and moves into the nucleus where it binds to specific DNA sequences. An alternatively spliced isoform that lacks amino acids (aa) 222‑231 (p65 delta ) does not bind DNA. Over the sequence used as an immunogen, human RelA p65 shares 96% and 98% aa identity with mouse and rat RelA p65, respectively. This portion includes one of eight potential ser/thr phosphorylation sites, two acetylation sites, and most of the Rel homology domain that interacts with I kappa B inhibitors.

 

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA

Entrez Gene IDs

5970 (Human)

Gene Symbol

RELA

UniProt

Q04206

Additional RelA/NFkB p65 Products

Product Documents for Human/Mouse/Rat RelA/NF kappa B p65 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human/Mouse/Rat RelA/NF kappa B p65 Antibody

For research use only

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

Customer Reviews for Human/Mouse/Rat RelA/NF kappa B p65 Antibody (2)

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Customer Images

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  • Human/Mouse/Rat RelA/NF kappa B p65 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Xenograft PD Sample
    Species: Human
    Verified Customer | Posted 10/24/2019
    Paired with AF5078 in ELISA.
    Human/Mouse/Rat RelA/NF kappa B p65 Antibody MAB5078
  • Human/Mouse RelA/NF kappa B p65 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Adenocarcinoma
    Species: Human
    Verified Customer | Posted 11/22/2016
    1x10^6 MCF7 cells were stained with 2.5 ug NFKBp65 antibody (red) and control antibody (blue). Blocked with 1% BSA, unfixed. Secondary antibody: PE-goat anti-mouse IgG with dilution 1:100.
    Human/Mouse/Rat RelA/NF kappa B p65 Antibody MAB5078

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Protocols

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