What is Odyssey blocking buffer?

Odyssey Blocking Buffer (PBS) is a ready-to-use formulation in phosphate-buffered saline that provides optimal blocking conditions for antibodies requiring PBS- based buffer systems. The blocker does not contain mammalian proteins.

What is the best blocking buffer for western blot?

Other considerations—Blocking buffers are often prepared in phosphate-buffered saline (PBS) or Tris- buffered saline (TBS). For detecting phosphorylated proteins, TBS is recommended because PBS may interfere with the antibody-antigen interaction.

What does blocking buffer do in Western blot?

The blocking buffer covers up or “blocks” these nonspecific protein-binding sites ensuring that most of your primary antibody binds only to the target protein (Figure 2). If you choose an effective blocking buffer, nonspecific binding can be reduced without interfering with antibody-antigen binding.

How do you make a BSA blocking buffer?

To make 100 mL of a 1% BSA blocking buffer, dissolve 1 g of BSA in 100 mL of TBST. The BSA blocking buffer recipe calculator enables the accurate preparation of BSA blocking solution whether you are making enough for a single experiment or for the entire lab.

Can you reuse BSA blocking buffer?

BSA functions as a protein stabilizer, allowing you to reuse antibodies longer when storing in the lab fridge and working with HRP. If you need to reuse your antibodies over an indefinite period of time between blots, you can make up the dilution in skim milk, without sodium azide, and freeze it at -20°C.

Why do you need methanol in transfer buffer?

The presence of methanol in the transfer buffer serves two main purposes: It promotes dissociation of SDS from the protein and dramatically improves adsorption of proteins onto membranes in the presence of SDS, although these effects may vary with proteins.

Why is SDS added to transfer buffer?

Adding SDS (up to 0.1%) to the transfer buffer increases the transfer efficiency of proteins, but reduces the amount of binding to the membrane. Therefore, if SDS is added to the transfer buffer, it is important to also include methanol (10–20%).