Hello, I'm having a lot of problems trying a paladin IF stain on skeletal muscle tissue. My negative controls seem to show more staining than my slides. For the negative control, I'm just omitting the primary antibody. I've tried increasing my blocking serum dilution and increasing the blocking time, but I haven't noticed a difference. Help!
Hi Mohamed, thanks for your reply. I'm using normal serum from the same species as the secondary antibody. When I use goat serum, it's at 5%, and when I use rabbit serum, I use a 1:20 dilution (per manufacturer's protocol). Originally I was blocking for 30 minutes but have increased that to an hour.
If I used BSA in my PBS rinses, what is a good concentration for this? I have BSA in liquid form but I have to verify its concentration.
Thanks!
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