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

Catalog # Availability Size / Price Qty
MAB5078
MAB5078-SP
Detection of Human and Mouse 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
Monoclonal Mouse IgG2B Clone # 532301
Purification
Protein A or G purified from hybridoma culture supernatant
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
2 µg/mL
See below
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
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below
Knockout Validated
RelA/NF kappa B p65 is specifically detected in HeLa humancervical epithelial carcinoma parental cell line but is not detectable inRelA/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 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 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 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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

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.

Immunocytochemistry RelA/NF?B p65 antibody in K562 Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Intracellular Staining by Flow Cytometry Detection of RelA antibody in HeLa Human Cell Line antibody by Flow Cytometry. View Larger

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.

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 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.

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 0.5 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 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
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. Radiotherapy Combined with PD-1 Inhibition Increases NK Cell Cytotoxicity towards Nasopharyngeal Carcinoma Cells
    Authors: A Makowska, N Lelabi, C Nothbaum, L Shen, P Busson, TTB Tran, M Eble, U Kontny
    Cells, 2021;10(9):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. The pro-inflammatory phenotype of the human non-classical monocyte subset is attributed to senescence
    Authors: SM Ong, E Hadadi, TM Dang, WH Yeap, CT Tan, TP Ng, A Larbi, SC Wong
    Cell Death Dis, 2018;9(3):266.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Intermittent Hypoxia Activates Duration-Dependent Protective and Injurious Mechanisms in Mouse Lung Endothelial Cells
    Authors: P Wohlrab, L Soto-Gonza, T Benesch, MP Winter, IM Lang, K Markstalle, V Tretter, KU Klein
    Front Physiol, 2018;9(0):1754.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

FAQs

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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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

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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

Paired with AF5078 in ELISA.


Human/Mouse RelA/NF kappa B p65 Antibody
By Anonymous on 11/22/2016
Application: Flow Cytometry Sample Tested: Adenocarcinoma Species: Human

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.