independent validation review for ha tag polyclonal antibody (stj90107)
TRANSCRIPT
Improving Quality And Reproducibility Through Antibody Validation
NEW INDEPENDENT VALIDATION RESULTS
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Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: HA-Tag Polyclonal Antibody (STJ90107)
Antibody: STJ90107. HA-tag Polyclonal AntibodyMethod for validation: Western BlotMaterials for validation: Hela cell line, Jurkat cell lineTreatment 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.Result of validation: target band was found in the material cell lines.
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Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: HA-Tag Polyclonal Antibody (STJ90107)
Primary antibody: Product Catalog No: STJ90107. HA-tag Polyclonal Antibody.100ul
Provider: St John's Laboratory
Dilution ratio: followed provider's recommendation
Secondary antibody:HRP goat anti-rabbit (provided by BPI)
Reference antibody:Actin (provided by BPI)
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Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: HA-Tag Polyclonal Antibody (STJ90107)
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).
PROTOCOL
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Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: HA-Tag Polyclonal Antibody (STJ90107)
Gel electrophoresis information： 10% polyacrylamide gel, constant voltage 160 V, 60 min.Transfer information： 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100 V 60 min.
No. AntigenLoading amount
Primary antibody
Primary antibody dilution
ratio
Secondaryantibody dilution
ratio
Target band KD
Visualization time
1 Hela lysate 10 ul St J90107. HA-tag
Polyclonal Antibody
1:1000
1:10000 68KD 34S
2 Jurkat lysate
10 ul 1:10000 68KD 34S
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Improving Quality And Reproducibility Through Antibody Validation
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