Q: I performed Western blot with NFkB p65 Antibody (NB100-2176) using brain and liver tissues extracts. My question: is it possible that NF-kB-Ikb complex could be detected by this antibody? Because I found 2 bands in the Western blot results, one is located at 65 and the other above 100 kD.

A: It possible that NB100-2176 could detect the entire NFkB complex in addition to just the p65 protein. If your samples were not completely reduced and denatured, they could have remained bound in your gel. The band above 100 kDa could also be an aggregate of NFkB. You may consider probing that same membrane for a different NFkB complex protein that you would expect to remain bound under these conditions. A negative signal in this case would tell you that it's an aggregate of NFkB, or maybe that it's a non-specific band. Did you run a negative control?

RelA/NFkB p65 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-2176

(3)

Citations (50)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
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Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Chemotaxis

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

View all RelA/NFkB p65 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide corresponding to the C-terminus of human NFkB p65.

Localization

Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I kappa B).

Specificity

Reacts with NFkB p65

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

69 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for RelA/NFkB p65 Antibody - BSA Free

Western Blot: RelA/NFkB p65 Antibody [NB100-2176]

Western Blot: RelA/NFkB p65 Antibody [NB100-2176] - Human glomerular endothelial cell lysates, time-response treatment with 10ng/ml TNF-alpha. This image was submitted via customer Review.

Immunohistochemistry-Paraffin: RelA/NFkB p65 Antibody [NB100-2176]

Immunohistochemistry-Paraffin: RelA/NFkB p65 Antibody [NB100-2176] - Staining of formalin-fixed, paraffin-embedded human DLBCL showing nuclear expression of NFkB p65 in the tumor cells.

Western Blot: RelA/NFkB p65 Antibody [NB100-2176]

Western Blot: RelA/NFkB p65 Antibody [NB100-2176] - Analysis of NFkB p65 expression in A431 (A) and HeLa (B) whole cell lysates.

Immunohistochemistry-Paraffin: Rabbit Polyclonal RelA/NFkB p65 Antibody [NB100-2176]

Immunohistochemistry-Paraffin: Rabbit Polyclonal RelA/NFkB p65 Antibody [NB100-2176] - RelA/NFkB p65 IHC (brown) in the mice liver tissue; bars, 200 μm. Image from a verified customer review.

Immunocytochemistry/Immunofluorescence: Rabbit Polyclonal RelA/NFkB p65 Antibody [NB100-2176]

Immunocytochemistry/Immunofluorescence: Rabbit Polyclonal RelA/NFkB p65 Antibody [NB100-2176] - Mice hepatocytes stained for RelA/NFkB p65. Image from a verified customer review.

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Applications for RelA/NFkB p65 Antibody - BSA Free

Application

Recommended Usage

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500

Western Blot

1:400-1:1000

Application Notes

Use in ICC/IF reported in scientific literature (PMID 27119753). Use in chromatin immunoprecipitation reported in scientific literature (PMID: 30411846).

Reviewed Applications

Read 3 reviews rated 5 using NB100-2176 in the following applications:

Immunofluorescence (1 Review)
Immunohistochemistry-Paraffin (1 Review)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 0.2% gelatin.

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: RelA/NFkB p65

p65 is a subunit of the nuclear factor kappa B. The transcription factor NFkB is widely recognized as a critical mediator of immune and inflammatory responses. In most cell types, NFkB is found in the cytoplasm where it is associated with an inhibitory protein known as IkB. An impressive variety of stimuli (tumor necrosis factor, interleukin 1, T cell activation signals, bacterial endotoxins, viral transforming proteins, certain growth factors and reactive oxygen intermediates) lead to the rapid nuclear accumulation of NFkB by the induced phosphorylation and subsequent degradation of IkB. In the nucleus, NFkB regulates genes encoding cytokines, cytokine receptors, cell adhesion molecules, proteins involved in coagulation and genes involved in cell growth control. Additionally, NFkB is thought to be an important transcriptional regulator for HIV. Growing evidence indicates that the dysregulation of NFkB may be key to a number of diseases including arthritis and other inflammatory diseases, Alzheimer's disease, atherosclerosis and cancer.

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA

Entrez Gene IDs

5970 (Human); 18033 (Mouse); 81736 (Rat)

Gene Symbol

RELA

UniProt

Q04206

Additional RelA/NFkB p65 Products

Product Documents for RelA/NFkB p65 Antibody - BSA Free

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Product Specific Notices for RelA/NFkB p65 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Apoptosis Transcription Factors and Regulators

C-type Lectin Receptors

IL-1 Family

Interferons

Natural Killer T (NKT) Cells

NFkB Pathway

NOD-like Receptors and the Inflammasome

PDGF Family

RIG-I-like Receptors

Toll-Like Receptors

Transcription Factors

Transcription Factors in the Akt Pathway

Citations for RelA/NFkB p65 Antibody - BSA Free

Customer Reviews for RelA/NFkB p65 Antibody - BSA Free (3)

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Customer Images

Showing 1 - 3 of 3 reviews Showing All

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RelA/NFkB p65 Antibody

Name: Armen Petrosyan

Application: Immunofluorescence

Sample Tested: Primary mouse hepatocytes

Species: Mouse

Verified Customer | Posted 11/19/2024

Mice hepatocytes stained for the NB100-2176
RelA/NFkB p65 Antibody

Name: Armen Petrosyan

Application: Immunohistochemistry-Paraffin

Sample Tested: Prostate tissue

Species: Mouse

Verified Customer | Posted 11/07/2024

NFkB p65 IHC (brown) in the mice liver tissue; bars, 200 μm.
RelA/NFkB p65 Antibody

Name: Anonymous

Application: Western Blot

Sample Tested: Endothelial and Whole cell lysates

Species: Human

Verified Customer | Posted 08/31/2017

Human glomerular endothelial cell lysates, time-response treatment with 10ng/ml TNF-alpha.

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for RelA/NFkB p65 Antibody - BSA Free

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Q: I performed Western blot with NFkB p65 Antibody (NB100-2176) using brain and liver tissues extracts. My question: is it possible that NF-kB-Ikb complex could be detected by this antibody? Because I found 2 bands in the Western blot results, one is located at 65 and the other above 100 kD.

A: It possible that NB100-2176 could detect the entire NFkB complex in addition to just the p65 protein. If your samples were not completely reduced and denatured, they could have remained bound in your gel. The band above 100 kDa could also be an aggregate of NFkB. You may consider probing that same membrane for a different NFkB complex protein that you would expect to remain bound under these conditions. A negative signal in this case would tell you that it's an aggregate of NFkB, or maybe that it's a non-specific band. Did you run a negative control?