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Actins are a group of proteins that play a role in different types of cell mortality. They are universally expressed in eukaryotic cells. Beta actin is one of six isoforms of actin that exist in human cells. It is also one of the two non-muscle cytoskeletal actins. The beta-actin antibody detects endogenous levels of beta-actin protein.
1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: beta-actin Monoclonal Antibody (STJ96941)
2. What was used? Primary antibody: STJ96941 beta-actin Antibody Provider: St John’s Laboratory Dilution ratio: 1:5000 Application: Western Blot Materials for validation: HEK293 cell lysates Click for product data sheet PDF
3. What Was The Protocol? Treatment of Materials When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors and equal volume of 2X loading buffer were added for 10 minutes in a water bath at 100?C. Gel Electrophoresis 12% Polyacrylamide gel with a constant voltage of 185V for 60 minutes. Transfer 0.45 um 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 overnight at 4¡ãC. Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% skimmed milk powder and left at room temperature for 1 hour. 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 a 1:5000 dilution ratio and left at room temperature for 1 hour. Membrane Wash Membrane was washed 3 times with 1X TBST. Visualization ECL visualization for 5 minutes (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: 12% Polyacrylamide gel with a constant voltage of 185V for 60 minutes. Transfer information: 0.45 um PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 minutes. No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 HEK293 lysate 10ul STJ96941 beta- actin antibody 1:5000 1:5000 ~43KD N/A
5. What were the results? beta-actin Antibody Predicted Band: 43 kDa Actual Band: 43 kDa
6. What did the customer think? Antibody Specificity: Antibody Rating: Testing results were provided independently by Viva Biotech Ltd. They stated ¡°Satisfied! The antibody works in accordance with expectation.¡±
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