Anyone use Superblock by Pierce? Do you use it according to Pierce's protocol (fill wells, immediately empty, repeat for a total of 3x)? This uses a lot of reagent.
I used to use SuperBlock for ELISA and Westerns. You can save time by following the wash 3 times directions, but you can also incubate longer (~15-20 minutes) and not do the three washes. Either way works fine.
Lately I've been using the Protein-Free Blocker from Pierce. It seems to have less background in most assays - probably because there is no protein to give nonspecific binding.
Here's the instructions for the Protein-Free Blocker:http://www.piercenet.com/files/1783as4.pdf
Have you used their Protein-Free block for western blotting? Does it reduce the background on the membrane?
I used to use it but it was an expensive alternative to milk or BSA and actually resulted in more background on Westerns. That said, it may be more useful for some antibodies than others.
Thanks for the info heehaw. I was afraid of that-higher background than what you normally get. I just recently got some non-animal protein blocker from another company and my background got worse-not better!! My primary was a rabbit monoclonal ab.