Question

Previous questions I do not need answered:

1) 1L Transfer Buffer: 25mM Tris, 192mM glycine, 10% methanol.
2)   100mls 1x TBS (10x TBS is supplied).
3)   400mls 1x TBS-T (Tris saline solution with 0.1% Tween). 10x TBS and Tween-20        will be provided.
4)   100mls Blocking buffer : 1x TBS-T/5%dry non fat milk

My questions:

1)   We will be using 15mls of a 1:5000 dilution of primary antibody in blocking buffer. Describe how you will prepare this reagent.
2) We will also be using 15mls of a 1:5000 dilution of secondary antibody in blocking    buffer. Describe how you will prepare this reagent.
3) What is the purpose of the blocking buffer?
4) What is the purpose of incubating the membrane with a primary and a secondary antibody?

Answer 1