Spreader-CDR system for efficient, reproducible and frugal western blot

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Title
Spreader-CDR system for efficient, reproducible and frugal western blot
Author(s)
Y R Cha; S Sajjad; K H Han; Tae-Sung Yoon; H S Shin; Sunghyun Kang
Bibliographic Citation
Electrophoresis, vol. 42, no. 14, pp. 1516-1520
Publication Year
2021
Abstract
Efficient antibody incubation is a vital step for successful western blot. During the incubation, a thin antibody-depleted layer is created around the blotting membrane, which limits antibody binding. Although the conventional batch shaking method is ineffective against it, this layer can be easily disrupted by cyclic draining and replenishing (CDR) of the antibody solution during membrane incubation. Previously, we introduced a closed and rotating cylindrical chamber as a tool to implement CDR for western blots (rCDR). A new open bucket-style chamber was devised for easier operation and the possibility of process automation. Instead of rotation as in rCDR, rocking it back and forth achieved the CDR antibody incubation (R-CDR). The chamber was then equipped with a spreader-rod to facilitate the uniform movement of the antibody solution across the membrane surface. Hence, it was named spreader CDR (S-CDR). Compared to the batch incubation method, both the S-CDR and R-CDR devices produced significantly enhanced signals and developed faster results. There were several additional benefits of using the spreader-rod, which included uniform antibody binding across the membrane, reduced usage of antibodies, and the ability to recover results even from mishandled, creased membranes. The S-CDR device ensures better blots and can be easily implemented in existing western blot protocols.
Keyword
Antibody incubationCDRS-CDRWestern blot
ISSN
0173-0835
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/elps.202000380
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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