Customer Review For NFKappaB-p65 Polyclonal Antibody (STJ94468)


Target information

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively.

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1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: NFKappaB-p65 Polyclonal Antibody (STJ94468)
2. WHAT WAS USED? Primary antibody: STJ94468. NFKappaB-p65 Polyclonal Antibody. 100ul Provider: St John’s Laboratory Dilution ratio: 1:1000 Application: Western Blot. Image: In HeLa cell line, there was a target band detected by STJ94468. NFKappaB-p65 Polyclonal Antibody.In A549 cell line, there were no target bands detected by STJ94468 NFKappaB-p65 Polyclonal Antibody.
3. WHAT WAS THE PROTOCOL? Treatment of materials: When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors (Sigma) and equal volume of 2X loading buffer were added for 10 min water bath at 100?C. Gel electrophoresis: 10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer: 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. Blocking: 1X TBST with 5% skimmed milk powder for 2 hr. Primary antibody probing: primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder, 4?C for overnight. Membrane wash: 1X TBST wash for 3 times. Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder with 1:10000 dilution ratio, for 1.5 hr. Membrane wash: 1X TBST wash for 3 times. Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that the visualization time was identical for the same antigen).
4. WHAT WAS THE PROTOCOL? Gel electrophoresis information£º10% polyacrylamide gel, constant voltage 160 V, 60 min. Transfer information£º0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min. No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 HeLa lysate 10 ug STJ94468. NFKappaB-p65 Polyclonal Antibody 1:1000 1:10000 65KD 30S 2 A549 lysate 10 ug 1:10000 65KD 30S
5. WHAT DID THE CUSTOMER THINK? Testing results were provided by BPI laboratory. They stated that ¡°In HeLa cell line, there was a target band detected by STJ94468. NFKappaB-p65 Polyclonal Antibody. In A549 cell line, there were no target bands detected by STJ94468 NFKappaB-p65 Polyclonal Antibody¡±.
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