Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract.
Lyse tissue samples in lysis buffer and measure total protein concentration to normalize loading amounts for subsequent gel electrophoresis.
Load denatured protein samples onto polyacrylamide gels and apply electric current to separate proteins by molecular weight through the gel matrix.
Electrophoretically transfer separated proteins from the gel onto a membrane where they remain immobilized for antibody-based detection.
Block non-specific binding sites on the membrane, then incubate with primary and secondary antibodies to visualize target protein bands.
Examine final blot images showing protein bands, interpret band intensity and position relative to molecular weight markers.
New protocols and pitfalls, in your inbox
A short email when we add notable lab videos and failure cases. No spam, unsubscribe anytime.