Antibody Customer Review for Acetyl-NFKB-p65 (K310) Polyclonal Antibody (STJ90133)

Target information

NFkB-P65 is a subunit of the NFkB transcription complex. This plays a significant role in inflammatory and immune responses. P65 has a weak DNA binding site which may contribute to binding the NFkB complex. In the cytoplasm IkB inhibits NFkB by interacting with P65.

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1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: Acetyl-NFKappaB-p65 (K310) Polyclonal Antibody (STJ90133)
2. WHAT WAS USED? Primary Antibody: Acetyl-NFKappaB-p65 (K310) Polyclonal Antibody (STJ90133) Provider: St John’s Laboratory Dilution ratio: 1:500 Application: Western Blot Materials for validation: HeLa cells
3. WHAT WAS THE PROTOCOL? Treatment of materials: Cells were lysed in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% Triton X-100, 1% Sodium deoxycholate, 1% SDS, supplemented with protease inhibitors (PMSF 1mM, Leupeptin 1ug/ml, Aprotinin 1ug/ml, Pepstatin 1ug/ml). Proteins were reduced and denatured by heating at 100¡æ for 5 min with subsequent cooling before use. Gel electrophoresis: 60ug/lane Transfer information: Nitocellulose membrane. Blocking: Blocked with 5% BSA for 0.5-1hr Primary Ab Application: Overnight at 4oC, 5% BSA in TBST. Secondary Ab Application: After washing with TBST 3 times membrane incubated secondary antibody for 1h at room temperature. Visualization: ECL kit
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