Chapter title |
A Novel Methodology for Stripping and Reprobing of Western Blots Originally Developed with Colorimetric Substrate TMB
|
---|---|
Chapter number | 35 |
Book title |
Detection of Blotted Proteins
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2718-0_35 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2717-3, 978-1-4939-2718-0
|
Authors |
Parmita Kar, Saurabh Kumar Agnihotri, Archana Sharma, Rekha Sachan, Madan Lal Bhatt, Monika Sachdev, Kar, Parmita, Agnihotri, Saurabh Kumar, Sharma, Archana, Sachan, Rekha, Bhatt, Madan Lal, Sachdev, Monika |
Abstract |
Protein blotting is a widely used analytical technique for detection of specific protein(s) in a given sample of tissue or cell homogenate or extract. Both chemiluminescence (CL) and colorimetric detections can be used for imaging protein blots in western blotting. Here we describe a methodology for stripping such western blots, already developed with the colorimetric substrate TMB on nitrocellulose membrane and reprobing the blot (after stripping) for the detection of a second protein through CL. The stripping procedure utilizes a stripping buffer consisting of β-mercaptoethanol, SDS, and Tris-HCl, while washing buffer consists of PBS added with 0.1 % Tween 20 and involves a series of steps facilitating accurate detection of the second protein in the stripped blot through CL. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 7 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 4 | 57% |
Student > Bachelor | 1 | 14% |
Other | 1 | 14% |
Unknown | 1 | 14% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 29% |
Chemistry | 2 | 29% |
Veterinary Science and Veterinary Medicine | 1 | 14% |
Medicine and Dentistry | 1 | 14% |
Unknown | 1 | 14% |