Hi,
I recall a few years back in a lab I used to work in that I used a 1:1 combination of 5% milk and 5% BSA in TBS-T for blocking and as an ab diluent for western blotting. It seemed to work well for particular phospho-proteins (e.g. Akt Thr308) from what I can remember).
Has anyone ever tried this and with any success? I find that while BSA on its own is best for phospho proteins, the blots are somewhat messy despite stringent washes.
Thanks in advance.
Dave
Blocking agent containing a mixture of non-fat dry milk and BSA can be used for certain antibodies. However, some optimization is needed. For most monoclonal antibodies and certain phospho-specific antibodies BSA alone may work better. Again, an optimization of blocking conditions may be needed.
Hi David J Walker ,
I used the following product for blocking (especially when I am probing against Phospho-proteins) and had much better results than using BSA alone. Worth a try...!
https://www.bio-rad-antibodies.com/reagent/block-ace-accessory-reagent-buf029.html?f=reagent
Thanks
Satish
In the case I could help you, I use this solution [10mM Tris-HCl pH 7.4, 5% fetal bovine serum (FBS), 1% Tween 20].
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