Posted 11 September 2011 - 11:43 PM
I need to perform a peptide blocking experiment for my antibody for western blot. I have never performed this and would appreciate any feedback.
I've read that the blocking peptides are used 10-100 times in excess of the antibody molarity? My antibody concentration is 2.5mg/ml and I use 1:250 dilution in TBST 5% milk for western blot. However I do not know how to calculate the amount of peptide to use.
My peptide concentration is 1mg/ml and has a calculated MW of 1876.55 Da. How much do I add to the TBST 5% milk?
Posted 12 September 2011 - 03:20 PM
IgG is normally about 150 kDa.
Posted 14 September 2011 - 05:36 AM