Purified anti-NF-κB p65 Antibody

Pricing & Availability
Clone
14G10A21 (See other available formats)
Regulatory Status
RUO
Other Names
Nuclear factor NF-kappa-B p65 subunit (NF-κB p65), Rel-A, transcription factor p65, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 (NFKB3)
Isotype
Mouse IgG2b, κ
1_14G10A21_PURE_NFkBp65_Antibody_2_WB_071217
Total lysates (15µg protein) from 293T and 293T/NF-κB p65 knockdown(KD) cells were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1µg/mL) purified anti-NF-κB p65 antibody, clone 14G10A21 (upper) or 1:3000 diluted anti-GAPDH (poly6314) antibody (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-NF-κB p65 antibody, and a donkey anti-rabbit IgG Antibody conjugated to HRP for GAPDH. Lane M: Molecular weight ladder, M* indicates longer exposure.
  • 1_14G10A21_PURE_NFkBp65_Antibody_2_WB_071217
    Total lysates (15µg protein) from 293T and 293T/NF-κB p65 knockdown(KD) cells were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 1:500 diluted (1µg/mL) purified anti-NF-κB p65 antibody, clone 14G10A21 (upper) or 1:3000 diluted anti-GAPDH (poly6314) antibody (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-NF-κB p65 antibody, and a donkey anti-rabbit IgG Antibody conjugated to HRP for GAPDH. Lane M: Molecular weight ladder, M* indicates longer exposure.
  • 2_14G10A21_PURE_NFkBp65_Antibody_1_WB_060716
    Hela cell extracts were resolved by electrophoresis, transferred to nitrocellulose, and probed with purified monoclonal anti-NF-κB p65 (clone 14G10A21) antibody. Proteins were visualized using an anti-mouse-IgG secondary conjugated to HRP and chemiluminescence detection.
  • 3_14G10A21_PURE_NFkBp65_Antibody_2_WB_060716
    Immunoprecipitation of NF-κB p65 from Hela cell extracts. Lane 1 is 5% input. Immunoprecipitation was performed using protein G resins only (lane 2), mouse IgG isotype control (lane 3), and anti-NF-κB p65 antibody (clone 14G10A21, lane 4). Western blot was performed using anti-NF-κB p65 antibody (clone 14G10A21).
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653001 25 µg $124.00
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653002 100 µg $265.00
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Description

NF-κB p65 is a member of REL-like domain containing protein family, which forms a NF-κB complex with the other family members: NF-κB1 (p105/p50) or NF-κB2 (p100/p52). The NF-κB complex is inactivated and held in the cytoplasm by the NF-κB inhibitor IκB. In response to activation stimuli, IκB kinases (IKKs) phosphorylates IκB, resulting in degradation of IκB and liberation of NF-κB complex. The activated NF-κB complex translocates to the nucleus and binds to κB sites in the DNA of their target genes.

Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Partial human NF-κB p65 recombinant protein (451-551 aa)
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.

Previous lots of this product may have been formulated with 0.1% or 0.05% NaN3 instead of 0.09% NaN3. For further information please contact BioLegend Technical Support or Customer Service.
Concentration
0.5 mg/ml
Storage & Handling
Upon receipt, store undiluted between 2°C and 8°C.
Application

WB - Quality tested
IP, KO/KD-WB - Verified

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1-1.0 µg per ml. For immunoprecipitation, the suggested use of this reagent is 2-10 μg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone is not recommended for ChIP (Chromatin Immunoprecipitation) assays (as determined by in-house testing).

Product Citations
  1. De Maeyer RPH, et al. 2020. Nat Immunol. 21:615. PubMed
RRID
AB_2561612 (BioLegend Cat. No. 653001)
AB_2561612 (BioLegend Cat. No. 653002)

Antigen Details

Structure
65 kD protein containing a Rel homology domain (RHD), an activation domain, and a 9aaTAD domain.
Distribution

The inactivated NF-κB complex containing p65 subunit is bound to IκB and is localized to cytoplasm. Upon activation, I-κB is phosphorylated and degraded. The activated NF-κB complex is in turn translocated to the nucleus as a transcription factor.

Function
NF-κB is a homodimeric or heterodimeric complex formed by the Rel-like domain-containing proteins. The most abundant form is p65 (RelA) - p50 (NF-κB1) heterodimer complex. The NF-κB complex is a ubiquitously expressed transcription factor which is involved in various biological functions, such as cell growth, tumorigenesis, differentiation, apoptosis, inflammation, and immune responses.
Interaction
Interacts with NF-κB1 (p105/p50) or NF-κB2 (p100/p52) to form heterodimeric NF-κB complex. Interacts with HDAC1, HDAC3, and CBP. Interaction with MEN1 inhibits transactivation activity of NF-κB complex.
Cell Type
B cells
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Immunology, Neuroscience, Neuroscience Cell Markers, Signal Transduction, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References

1. Li Z, et al. 1997. Mol. Cell. Biol. 17:6184.
2. Saccani S, et al. 2004. J. Exp. Med. 200:107.
3. Nolan GP, et al. 1991. Cell 64:961.
4. Chen LF, et al. 2001. Science 293:1653.
5. Hansen SK, et al. 1994. Mol. Cell. Biol. 14:2593.
6. Chapman NR, et al. 2002. Biochem J. 366:459.

Gene ID
4790 View all products for this Gene ID
UniProt
View information about NF-kappaB p65 on UniProt.org
Go To Top Version: 3    Revision Date: 02/08/2024

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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