Western Blotting: Difference between revisions
added links to buffers |
m →Protocol: Changed wash times |
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#Block with 2% BSA in [[ TBST ]] or 5% skim milk powder in TBST for >1h | #Block with 2% BSA in [[ TBST ]] or 5% skim milk powder in TBST for >1h | ||
#Incubate with primary antibody (check for dilution) in 1 mg/mL BSA for >1h | #Incubate with primary antibody (check for dilution) in 1 mg/mL BSA for >1h | ||
#Wash blot every | #Wash blot every 5 minutes for 15 min with TBST. | ||
#Incubate with appropriate secondary antibody (10 000X) for 45min-1h | #Incubate with appropriate secondary antibody (10 000X) for 45min-1h | ||
#Wash blot every | #Wash blot every 5 minutes for 15 min with TBST. | ||
#Rinse once or twice with double distilled water | #Rinse once or twice with double distilled water | ||
#Prepare ECL by mixing one volume of the black bottle solution with one volume of the white bottle solution (be careful not to cross-contaminate ECL bottles with wrong solution). | #Prepare ECL by mixing one volume of the black bottle solution with one volume of the white bottle solution (be careful not to cross-contaminate ECL bottles with wrong solution). | ||