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Inhibitor of nuclear factor Kappa-B Kinase (IKK) is a Serine kinase which is essential for the NFkB pathway. IKK-alpha and -beta are subunits of the IKK complex. The kinase ability phosphorylates inhibitors of NFkB to allow free NFkB to translocate to the nucleus, where it activates transcription of genes involved in immune and inflammatory responses.
1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: Phospho-IKKalpha/beta (S176/177) Polyclonal Antibody (STJ90301)
2. WHAT WAS USED? Primary Antibody: Phospho-IKKalpha/beta (S176/177) Polyclonal Antibody (STJ90301) Provider: St John’s Laboratory Dilution ratio: 1:500 Application: Western Blot Materials for validation: NIH3T3 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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