Detailed Explanation
Laboratory technique for detecting specific peptides or proteins in a sample. The process has four steps: (1) proteins are separated by size using gel electrophoresis (SDS-PAGE), (2) separated proteins are transferred ('blotted') from the gel onto a membrane (nitrocellulose or PVDF), (3) the membrane is probed with a primary antibody specific to the target peptide, followed by an enzyme-linked secondary antibody, (4) a substrate reaction produces a visible band at the position of the target.
The name 'Western' follows the naming convention of Southern blot (DNA, by Edwin Southern) and Northern blot (RNA) — Western blot was named as a directional joke by W. Neal Burnette in 1981. Western blotting is a standard validation method in peptide and protein research.
Key Facts
- Laboratory technique for detecting specific peptides or proteins in a sample.
- The process has four steps: (1) proteins are separated by size using gel electrophoresis (SDS-PAGE), (2) separated proteins are transferred ('blotted') from the gel onto a membrane (nitrocellulose or PVDF), (3) the membrane is probed with a primary antibody specific to the target peptide, followed by an enzyme-linked secondary antibody, (4) a substrate reaction produces a visible band at the position of the target.
- The name 'Western' follows the naming convention of Southern blot (DNA, by Edwin Southern) and Northern blot (RNA) — Western blot was named as a directional joke by W.
- Western blotting is a standard validation method in peptide and protein research.
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