A systematic approach to quantitative Western blot analysis.

Authors :: Pillai-Kastoori L
Schutz-Geschwender AR
Harford JA
Source :: Analytical biochemistry [Anal Biochem] 2020 Mar 15; Vol. 593, pp. 113608. Date of Electronic Publication: 2020 Jan 31.
Publication Year :: 2020
Abstract: 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.<br />Competing Interests: Declaration of competing interest L. Pillai-Kastoori, A. Schutz-Geschwender, and Jeff A. Harford are employees of LI-COR Biosciences.<br /> (Copyright © 2020 LI-COR BIOSCIENCES, LINCOLN, NEBRASKA 68504. Published by Elsevier Inc. All rights reserved.)