RelA/NFkB p65 [p Ser529] Antibody
Novus Biologicals, part of Bio-Techne | Catalog # 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.
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] - 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] - 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.
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
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
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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