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] - 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] - 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.

• Reactivity: Hu
• Applications: WB, Simple Western, ELISA, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

RelA/NFkB p65 [p Ser529] Antibody Summary

• 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.
• 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)
• 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.
• Isotype: Serum
• Clonality: Polyclonal
• Host: Rabbit
• Gene: RELA
• Purity: Delipidation and Defibrination

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000-1:25000
  • Immunohistochemistry 1:200-1:1000
  • Simple Western
  • 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.
• Control Peptide:

  RelA/NFkB p65 Protein (NBP1-97000)
  RelA/NFkB p65 Protein (NBP1-97001)

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.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Antiserum
• Preservative: 0.1% Sodium Azide
• Purity: Delipidation and Defibrination

Alternate Names for RelA/NFkB p65 [p Ser529] Antibody

• MGC131774
• NF kB p65 phospho
• nf kb p65
• NFkB p65 phospho
• NF-kB p65 phospho
• 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
• phospho NF kB p65
• phospho NFkB p65
• phospho NF-kB p65
• 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

Background

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.

Limitations

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.

Publications for RelA/NFkB p65 Antibody (NBP1-77808)(6)

Publications using NBP1-77808

• Lu SM, Tremblay MÈ, King IL et al. HIV-1 Tat-induced microgliosis and synaptic damage via interactions between peripheral and central myeloid cells. PLoS One 2011-01-01 [PMID: 21912650]
• Shakibaei M, Buhrmann C, Mobasheri A et al. Anti-inflammatory and anti-catabolic effects of TENDOACTIVE on human tenocytes in vitro. Histol Histopathol 2011-09-01 [PMID: 21751149]
• Castier Y, Ramkhelawon B, Riou S et al. Role of NF-kappaB in flow-induced vascular remodeling. Antioxid Redox Signal 2009-07-01 [PMID: 19320561]
• Zhao L, Lee JY, Hwang DH et al. The phosphatidylinositol 3-kinase/Akt pathway negatively regulates Nod2-mediated NF-kappaB pathway. Biochem Pharmacol 2008-04-01 [PMID: 18243161]
• Shakibaei M, John T, Schulze-Tanzil G et al. Suppression of NF-kappaB activation by curcumin leads to inhibition of expression of cyclo-oxygenase-2 and matrix metalloproteinase-9 in human articular chondrocytes: Implications for the treatment of osteoarthritis. Biochem Pharmacol. 2007-05-01 [PMID: 17291458]
• Ashikawa K, Majumdar S, Banerjee S et al. Piceatannol inhibits TNF-induced NF-kappaB activation and NF-kappaB-mediated gene expression through suppression of IkappaBalpha kinase and p65 phosphorylation. J Immunol. 2002-12-01 [PMID: 12444159]

Bioinformatics

• Gene Symbol: RELA
• Entrez:
  • 5970(Human)
• Uniprot:
  • Q04206()
  • Q04206()
  • Q04206()
  • Q04206()
  • Q04206()