This phosphospecific polyclonal antibody reacts with p65 (RelA) pS536 and shows minimal reactivity by western blot with nonphosphorylated p65 (RelA) and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide.
交叉反应 (详细)
Species reactivity (expected):Mouse, Rat (100 %)Species reactivity (tested):Human.
纯化方法
Immunoaffinity chromatography using phospho-peptide coupled to agarose beads followed by solid phase adsorption against nonphospho-peptide
免疫原
Synthetic peptide corresponding to residues surrounding S536 of human p65 (RelA) protein
ELISA: 1: 1,000 - 1: 6,000. Western blot: 1: 200 - 1: 2,000 (a band approximately 65 kDa in size corresponding tophosphorylated p65 (RelA) protein is expected in the appropriate cell lysate or extract). Immunohistochemistry on paraffin sections: 5 μg/mL. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
限制
仅限研究用
浓度
0.9 mg/mL (by UV absorbance at 280 nm)
缓冲液
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01 % (w/v) Sodium Azide
储存液
Sodium azide
注意事项
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
注意事项
Avoid repeated freezing and thawing. Should this product contain a precipitate we recommend microcentrifugation before use.
储存条件
4 °C/-20 °C
储存方法
Store the antibody at 2 - 8 °C up to one month or for longer (in aliquots) at -20 °C or below.
NFkappaB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells. It was subsequently found in non-B cells in an inactive cytoplasmic form consisting of NFkappaB bound to IkappaB. NFkappaB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NFKB1) subunits. Other identified subunits include p52 (NFKB2), cRel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkappaB subunit p65, similar to p50/p65 heterodimers. Lowlevels of p52 and p50 homodimers can also exist in cells. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkappaB-alpha. IkappaB-alpha binds to the p65 subunit, preventing nuclear localization, and DNA binding. Activators mediate a rapid phosphorylation of IkappaB by IkappaB kinase (IKK), which results in subsequent ubiquitination and proteolytic degradation. NFkappaB is then transported to the nucleus, where it activates transcription of target genes through binding to NFkappaB target sequences within the promoter. The HTLV-I protein Tax can induce constitutive NFkappaB activation through phosphorylation of both IkappaB-alpha and IkappaB-beta. The transforming protein Tax inhibits p53 transcriptional activity through the NFkappaB signaling pathway, specifically via the p65 (RelA) subunit. The inhibition of p53 activity is dependent upon phosphorylation of p65 (RelA) at S536 by the upstream kinase IKKbeta.Synonyms: NF kappa B p65, NFKB3, NFkB p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, Rel A, Transcription factor p65