Dealing with Large Sample Sizes: Comparison of a New One Spot Dot Blot Method to Western Blot

Putra, Sulistyo Emantoko Dwi and Tsuprykov, Oleg and Websky, Karoline Von and Ritter, Teresa and Reichetzeder, Christoph and Hocher, Berthold (2014) Dealing with Large Sample Sizes: Comparison of a New One Spot Dot Blot Method to Western Blot. Clinical laboratory, 60 (11). pp. 1-7. ISSN 14336510

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Abstract

Background: Western blot is the gold standard method to determine individual protein expression levels. However, western blot is technically difficult to perform in large sample sizes because it is a time consuming and labor intensive process. Dot blot is often used instead when dealing with large sample sizes, but the main disad- vantage of the existing dot blot techniques, is the absence of signal normalization to a housekeeping protein. Methods: In this study we established a one dot two development signals (ODTDS) dot blot method employing two different signal development systems. The first signal from the protein of interest was detected by horseradish peroxidase (HRP). The second signal, detecting the housekeeping protein, was obtained by using alkaline phos- phatase (AP). Results: Inter-assay results variations within ODTDS dot blot and western blot and intra-assay variations between both methods were low (1.04 - 5.71%) as assessed by coefficient of variation. Conclusions: ODTDS dot blot technique can be used instead of western blot when dealing with large sample sizes without a reduction in results accuracy.

Item Type:	Article
Uncontrolled Keywords:	one dot two development signals (ODTDS) dot blot, western blot, protein quantification, large sample size studies, comparison
Subjects:	T Technology > T Technology (General)
Divisions:	Faculty of Technobiology > Department of Biology
Depositing User:	Ester Sri W. 196039
Date Deposited:	23 Mar 2021 08:00
Last Modified:	10 Nov 2021 06:40
URI:	http://repository.ubaya.ac.id/id/eprint/39065

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