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RelA/NFkB p65 [p Ser529] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-77808

Novus Biologicals, part of Bio-Techne
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NBP1-77808

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

ELISA, Immunohistochemistry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit Serum

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

Immunogen

RelA/NFkB p65 peptide corresponding to a region near phospho Serine 529 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH). (Uniprot: Q04206)

Reactivity Notes

This phospho specific polyclonal antibody is specific for phosphorylated pS529 human RelA/NFkB p65. Reactivity with non-phosphorylated RelA/NFkB p65 is minimal. Cross reactivity with pS529 phosphorylated RelA/NFkB p65 from mouse, rat or other species has not been determined.

Modification

p Ser529

Specificity

This phospho specific polyclonal antibody is specific for phosphorylated pS529 human RelA/NFkB p65. Reactivity with non-phosphorylated RelA/NFkB p65 is minimal. Cross reactivity with pS529 phosphorylated RelA/NFkB p65 from mouse, rat or other species has not been determined.

Clonality

Polyclonal

Host

Rabbit

Isotype

Serum

Description

This antibody was prepared from monospecific antiserum by delipidation and defibrination

Store this antibody at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for RelA/NFkB p65 [p Ser529] Antibody

Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808] - TNF Induces phosphorylation of p65 in KBM-5 cells. Nuclear protein lysates prepared after 0, 5, 10, 15, 30 and 60 minutes of 0.1 nM TNF treatment of KBM-5 cells shows inducible phosphorylation using phospho specific polyclonal anti-human pS529 p65. Anti-beta-actin staining confirms loading of equivalent amounts of protein. HRP conjugated Gt-anti-Rabbit IgG used to develop blot using a chemiluminescent detection method. Other detection methods will yield similar results.
Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808]

Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808]

Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808] - pS529 shows phospho p65 staining in carcinoma cells. western blot of total protein lysates from various human head and neck tumors shows phospho p65 staining in tumor cell lines using phospho specific polyclonal anti-human pS529 p65. Lanes 1-6 contain protein lysates from human squamal carcinoma cell lines. Lane 7 is a protein lysate from a primary culture of human keratinocytes. Lane 8 contains protein lysate from ATCC SCC9 cells (also a head and neck squamal carcinoma). Lane 9 contains lysate from EGF-induced human derived A431 cells. Lane 10 (not shown) contains a molecular weight standard. Concurrent staining with anti-beta microtubulin (not shown) was used to confirm equal protein loading in all lanes. HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method.
Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808]

Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808]

Western Blot: RelA/NFkB p65 [p Ser529] Antibody [NBP1-77808] - TNF Induces phosphorylation of p65 in KBM-5 cells. Cytoplasmic and nuclear protein lysates prepared after 0, 5, 10, 15, 30 and 60 minutes of 0.1 nM TNF treatment of KBM-5 cells shows inducible phosphorylation using phospho specific polyclonal anti-human pS276 p65. Pan reactive anti p65 (NBP1-77808) was used a control to show the presence of total p65 in both the cytoplasmic and nuclear extracts. Phosphorylation of p65 occurs after approximately 10 min of TNF exposure. Migration of phosphorylated p65 into the nucleus occurs within a similar time frame. HRP conjugated Gt-anti-Rabbit IgG was used to develop the western blot using a chemi-luminescent detection method.

Applications for RelA/NFkB p65 [p Ser529] Antibody

Application
Recommended Usage

ELISA

1:5000-1:25000

Immunohistochemistry

1:200-1:1000

Western Blot

1:500-1:3000
Application Notes
This product reacts human pS529 p65 and shows minimal reactivity by western blot with non-phosphorylated p65 and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.

Simple Western Antibody Database for Simple Western validation: tested in HeLa-/+TNF, HeLa-/+TPA, nuclear extract; separated by charge; antibody dilution of 1:50.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Delipidation and Defibrination

Formulation

Antiserum

Preservative

0.1% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: RelA/NFkB p65

Nuclear factor B (NF-B) encompasses an important family of inducible transcriptional activators that regulate a wide variety of cellular and viral genes. The family members include p50, p52, p65 (RelA), c-Rel, and RelB. The Nf-kB p65 subunit, similar to two others in the B family, RelB and c-Rel, contains two transactivation domains in the C-terminal region of the protein. Association with inhibitory proteins of the I B family, retains NF- B in the cytoplasm. Degradation of IB proteins exposes the nuclear localization sequence (NLS), leading to nuclear translocation and subsequent binding of NF-B to DNA. In addition to the nuclear translocation and DNA binding, Nf-kB's transcriptional activity is also regulated by coactivators and corepressors in the nucleus. Interaction of p65 with coactivators; CREB-binding protein and p300, increases its transcriptional activity.

Long Name

v-rel Reticuloendotheliosis Viral Oncogene Homolog A

Alternate Names

NFkB p65, NFKB3, p65RelA, NF kB p65 phospho, NFkB p65 phospho, NF-kB p65 phospho, phospho NF kB p65, phospho NFkB p65, phospho NF-kB p65

Entrez Gene IDs

5970 (Human)

Gene Symbol

RELA

UniProt

Additional RelA/NFkB p65 Products

Product Documents for RelA/NFkB p65 [p Ser529] Antibody

Certificate of Analysis

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

Product Specific Notices for RelA/NFkB p65 [p Ser529] Antibody

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.

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