Mastering the Western Blot Technique

Western Blot Technique

• The western blot technique is used to detect specific proteins in a sample of tissue homogenate or extract.
• The three elements used in the western blot technique are:
  • Electrotransfer of proteins to a blotting membrane
  • Immunodetection with primary and secondary antibodies
  • Chemiluminescent or chromogenic detection of the antigen-antibody complex

Role of Secondary Antibody

• The purpose of the secondary antibody in the western blot technique is to bind to the primary antibody, amplifying the signal and allowing for detection.

Detection of Biomarkers

• Western blot is a method commonly used to detect biomarkers in the diagnosis of neurological and oncological illnesses.

Multidrug Resistance (MDR) in Cancer Therapy

• Multidrug resistance (MDR) plays a significant role in cancer therapy, making it challenging to treat cancer patients.

Creutzfeldt-Jakob Disease Diagnosis

• Western blotting analysis of the isoform pattern of 14-3-3 proteins in cerebral fluid can identify Creutzfeldt-Jakob disease.

Unique Feature of Western Blotting

• The main feature that sets western blotting apart from other epitope mapping techniques is its ability to identify specific protein isoforms.

History of Western Blot

• Western blot was independently invented in 1979 by Jaime Renart, Jakob Reiser, and George Stark at Stanford University, and by Harry Towbin, Theophil Staehelin, and Julian Gordon at the Friedrich Miescher Institute in Basel, Switzerland.

Prevalence of Western Blot in Research

• Approximately 10% of protein-related publications apply western blots.

Medical Diagnostics

• The western blot test is employed to detect anti-HIV antibody in a human serum sample.
• Western blot is used as the definitive test to confirm Lyme disease.