Rapid and efficient western blot assay by rotational cyclic draining and replenishing procedure = 연속 배수 및 보충 방식을 이용한 신속하고 효율적인 웨스턴 블롯 분석

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Title
Rapid and efficient western blot assay by rotational cyclic draining and replenishing procedure = 연속 배수 및 보충 방식을 이용한 신속하고 효율적인 웨스턴 블롯 분석
Author(s)
Saba Sajjad; M T Do; H S Shin; Tae-Sung YoonSunghyun Kang
Bibliographic Citation
Electrophoresis, vol. 39, no. 23, pp. 2974-2978
Publication Year
2018
Abstract
Western blot is a principal technique for the detection of specific proteins in various biology disciplines. The antibody incubation is an imperative step in western blot. Antibody incubation by mild shaking on a rocker is generally used to facilitate mixing of antibodies. However, mild shaking is an inefficient process to remove antibody depletion layer on blotting membrane and requires hours of incubation time to achieve antibody binding to target proteins. We propose an alternative method of cyclic draining and replenishing (CDR) incubation of antibody solution using a rotational incubation chamber. The study demonstrated that rotational CDR incubation could shorten antibody incubation time with enhanced sensitivity. Moreover, rotational CDR incubation could achieve a stronger antibody binding with lower antibody concentration when compared with batch incubation. In addition, rotational CDR incubation significantly improved the detection of low abundance proteins. This simple modification in western blot procedure makes it rapid, sensitive, and cost-efficient.
Keyword
Antibody incubationRotational CDRWestern blot
ISSN
0173-0835
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/elps.201800195
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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