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A systematic approach to quantitative Western blot analysis

2020; Elsevier BV; Volume: 593; Linguagem: Inglês

10.1016/j.ab.2020.113608

1096-0309

Lakshmi Pillai‐Kastoori, Amy Schutz‐Geschwender, Jeff A. Harford,

Enzyme Structure and Function

Attaining true quantitative data from WB requires that all the players involved in the procedure are quality controlled including the user. Appropriate protein extraction method, electrophoresis, and transfer of proteins, immunodetection of blotted protein by antibodies, and the ultimate step of imaging and analyzing the data is nothing short of a symphony. Like with any other technology in life-sciences research, Western blotting can produce erroneous and irreproducible data. We provide a systematic approach to generate quantitative data from Western blot experiments that incorporates critical validation steps to identify and minimize sources of error and variability throughout the Western blot process.