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Western Blot Antibody Customer Review for Anti-RelB Polyclonal Antibody (STJ95410)

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The RelB protein is a transcription factor encoded by the RELB gene in humans. It is known to interact with NFKB2. The RelB polyclonal antibody detects endogenous levels of RelB protein.

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Western Blot Antibody Customer Review for Anti-RelB Polyclonal Antibody (STJ95410)

  1. 1. Result “Specific band in HeLa extracts. Not Specific in NRK extracts.” - A. Diaz, Centro Nacional de Biotecnologia CNB CSIC. Cell Line: HeLa, NRK Method of validation: Western Blot Primary Antibody: STJ95410 Anti-RelB Antibody Secondary Antibody: HRP goat anti-rabbit Dilution ratio: 1:1000 Protocol Treatment of materials: When cells were cultured to 80% plating density, lysis buffer with triton X-100 were added, soluble and insoluble frac- tions were separated by centrifugation and sample buffer was added and then boiled. Gel electrophoresis: 10% polyacrylamide gel, constant voltage of 160V for 60 minutes. Transfer: 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes. Blocking: 1X TBST with 5% skimmed milk powder was used and left for 1 hour. Primary antibody probing: Primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder at 4˚C and left overnight. Membrane wash: Membrane was washed 3 times with 1X TBST. 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 hour. Membrane wash: Membrane was washed 3 times with 1X TBST. Visualization: : ECL visualization for 5 - 10 minutes. Antibody Customer Review: STJ95410 Anti-RelB Antibody Antibody Specificity: Antibody Rating: Gel electrophoresis information: 10% polyacrylamide gel, constant voltage of 160V for 60 minutes. Transfer information: 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes. No. Antigen Loading amount Primary anti- body Primary antibody dilution ratio Secondary antibody dilu- tion ratio Target band KD Visualization time 1 As said in image 10µg STJ95410 Anti- RelB Antibody 1:1000 1:10000 ~72/60KD N/A

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