Immunoblot

Definition

An immunoblot, also known as a Western blot, is a laboratory method used to detect specific proteins in a sample using antibody binding. It is commonly used to analyze the immune response and protein expression.

5 Must Know Facts For Your Next Test

1. Immunoblots involve separating proteins by gel electrophoresis before transferring them to a membrane.
2. Primary antibodies are used to specifically bind to the target protein on the membrane.
3. Secondary antibodies, which are often conjugated with enzymes or fluorescent markers, bind to primary antibodies for detection.
4. Commonly used detection methods include chemiluminescence and colorimetric assays.
5. Blocking agents like non-fat milk or BSA are used to prevent non-specific binding of antibodies.

Review Questions

• What is the purpose of using primary and secondary antibodies in an immunoblot?
• Can you name two common methods of detecting proteins in an immunoblot?
• Why are blocking agents necessary in an immunoblot procedure?

Related terms

ELISA:

A plate-based assay technique for detecting and quantifying soluble substances such as peptides, proteins, antibodies, and hormones.

Gel Electrophoresis:

A method for separation and analysis of macromolecules (DNA, RNA, proteins) based on their size and charge.

Antibody:

A Y-shaped protein produced by B-cells that binds specifically to antigens.