cheminuminescent protein ladder for western blot

Recent advancements in blocking agents have helped make this step faster and more efficient. Proteins in this blocking solution occupy the empty spaces on the membrane to help prevent detection antibodies in the next step from sticking to the membrane. To do this the membrane should first be incubated with a protein containing solution such as nonfat dry milk or a purified protein like bovine serum albumin, or BSA. This is a multi-step process, and the first step is minimizing nonspecific binding by using a blocking agent. Once complete, the efficiency of the transfer can be confirmed by staining the membrane with a reversible stain. However, new technologies have been developed that can cut the transfer time down to as little as 7 minutes. Using a traditional apparatus, the protein transfer process can take 30 to 60 minutes to complete.

Once the proteins have been separated in the gel, they must be transferred to a solid support membrane, which facilitates protein detection through the use of antibodies specific to the target protein. In this video, we will cover the 3 major steps in generating a western blot: Separate, Transfer, and Detect.Īfter protein samples have been prepared, the mixture is separated by molecular weight using protein gel electrophoresis, which is a standard laboratory technique by which charged protein molecules are transported through a matrix by an electrical field. Western Blotting is the most widely used technique for protein detection.

Cheminuminescent protein ladder for western blot