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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

Authors 

Saba SajjadM T DoH S ShinTae-Sung YoonSunghyun Kang

Publisher 

Wiley

Issue Date 

2018

Citation 

Electrophoresis

Keywords 

Antibody incubationRotational CDRWestern blot

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.

ISSN 

0173-0835

Link 

http://dx.doi.org/10.1002/elps.201800195

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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