What does blocking with BSA do?

BSA blocking is a routine practice among clinicians and researchers working on immunoassays throughout the world. The primary role of BSA is to prevent the non-specific binding by blocking the leftover spaces over solid surface after immobilization of a capture biomolecule.

What does blocking do in IHC?

Blocking with sera or a protein blocking reagent prevents non-specific binding of antibodies to tissue or to Fc receptors. Theoretically, any protein that does not bind to the target antigen can be used for blocking.

Which BSA is used for blocking?

Bovine serum albumin (BSA) blocking buffer is ideal for saturating excess protein-binding sites on membranes and microplates for Western blotting and ELISA applications, respectively. Typically, 1-3% BSA is sufficient for most applications.

How do I block cells with BSA?

Block with 5 % normal goat serum/PBS or 1 % BSA/PBS for 45 minutes (no washing required). Dilute the primary antibody in blocking solution and apply it for 2 h (or overnight at 4 °C). Wash 4 × thoroughly to remove unbound primary antibody.

What is BSA blocking buffer?

Blocker BSA (10X) is purified 10% solution of bovine serum albumin for blocking in western blotting, ELISA, IHC and nucleic acid detection methods. Blocker BSA is a 10% (w/v) solution of high-quality BSA that is useful for saturating excess protein-binding sites on membranes and microplates in immunoassays.

Why is blocking done?

Blocking is used to remove the effects of a few of the most important nuisance variables. Randomization is then used to reduce the contaminating effects of the remaining nuisance variables. For important nuisance variables, blocking will yield higher significance in the variables of interest than randomizing.

What percentage is BSA for blocking?

Blocker BSA (10X) is a purified 10% solution of bovine serum albumin for blocking in western blotting, ELISA, IHC, and nucleic acid detection methods. Blocker BSA is a 10% (w/v) solution of high-quality BSA that is useful for saturating excess protein-binding sites on membranes and microplates in immunoassays.

How much is BSA for blocking?

Why is BSA added to buffers?

1 BSA addition increases significantly the total pro- tein concentration, thus stabilizing other biological reagents by making them less susceptible to proteolysis or simply by decreas- ing water activity.

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.