hh Human/Mouse RelA/NF kappa B p65 Antibody MAB5078: R&D Systems

Human/Mouse RelA/NF kappa B p65 Antibody

(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse 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 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
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 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.
Immunocytochemistry
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 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 in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human/Mouse 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 kappa B p65Specificity 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 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 HeLacell 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • 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
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Flow Cytometry.

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Flow Cytometry Human/Mouse RelA/NF kappa B p65 Antibody MAB5078
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 Flow Human Anonymous 11/22/2016
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Flow Cytometry Human/Mouse RelA/NF kappa B p65 Antibody MAB5078
Flow Cytometry: Human/Mouse RelA/NF kappa B p65 Antibody [MAB5078]

Summary

ApplicationFlow Cytometry
Sample TestedAdenocarcinoma
SpeciesHuman

Other Experimental Details

Other Experimental Details1x10^6 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.

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