A rapid method for large sample number extraction of proteins from mammalian cells

Finding the best method of cell lysis and extraction of protein is the key step in detection and identification of proteins in all applications of proteomics. The methods currently used for protein extraction from mammalian cells are usually laborious, expensive or insufficiently reliable. R...

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Main Authors:	Packeer Mohamed, Vasila, Hashim, Yumi Zuhanis Has-Yun, Amid, Azura, Mohd Salleh, Hamzah, Yusof, Faridah, Mel, Maizirwan
Format:	Article
Language:	English
Published:	Tissue Engineering and Regenerative Medicine Society of Malaysia 2012
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/27821/ http://irep.iium.edu.my/27821/ http://irep.iium.edu.my/27821/1/Paper_-_Packeer-Mohamed_et_al_2012_%28RegRes%29.pdf

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recordtype	eprints
spelling	iium-278212013-02-14T00:11:25Z http://irep.iium.edu.my/27821/ A rapid method for large sample number extraction of proteins from mammalian cells Packeer Mohamed, Vasila Hashim, Yumi Zuhanis Has-Yun Amid, Azura Mohd Salleh, Hamzah Yusof, Faridah Mel, Maizirwan Q Science (General) Finding the best method of cell lysis and extraction of protein is the key step in detection and identification of proteins in all applications of proteomics. The methods currently used for protein extraction from mammalian cells are usually laborious, expensive or insufficiently reliable. Recently, researchers working with mammalian cells conduct protein extraction widely and frequently. This frequent dealing with a large number of samples becomes tedious work. Even though there are many commercial lysis buffers on the market, they are usually expensive. Therefore, in this study, protein extraction lysis buffer was designed to provide highly efficient total protein extraction from cultured mammalian cells. The simple and unique composition of this lysis buffer containing sodium dodecyl sulphate (SDS) dissolves cell membranes and extracts total cellular protein in only 45 minutes. The novel extraction method and lysis buffer composition was formulated using classical optimization or one factor at a time (OFAT) study. In this method, cell pellet from mammalian cells (CHO-K1) were subjected to different lysis buffer treatment procedure and then incubated at room temperature. The SDS-PAGE was performed to confirm the proteins extracted. The method was extended to bacteria (Escherichia coli, BL21-AI), yeast (Pichia pastoris) and repeated for mammalian (CHOK1) for confirmation. It yields complete extraction for bacteria and mammalian cells. As such, this lysis buffer when used for preparing protein extracts from bacteria and mammalian cells offers rapid, reliable and cheap protocol which is applicable to routine laboratory analysis especially for protein estimation and SDS-PAGE. Tissue Engineering and Regenerative Medicine Society of Malaysia 2012-12 Article PeerReviewed application/pdf en http://irep.iium.edu.my/27821/1/Paper_-_Packeer-Mohamed_et_al_2012_%28RegRes%29.pdf Packeer Mohamed, Vasila and Hashim, Yumi Zuhanis Has-Yun and Amid, Azura and Mohd Salleh, Hamzah and Yusof, Faridah and Mel, Maizirwan (2012) A rapid method for large sample number extraction of proteins from mammalian cells. Regenerative Research, 1 (2). pp. 49-55. ISSN 2232-0822 (Online) http://regres.tesma.org.my/pdf/RR-151012-006%288%29.pdf
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General)
spellingShingle	Q Science (General) Packeer Mohamed, Vasila Hashim, Yumi Zuhanis Has-Yun Amid, Azura Mohd Salleh, Hamzah Yusof, Faridah Mel, Maizirwan A rapid method for large sample number extraction of proteins from mammalian cells
description	Finding the best method of cell lysis and extraction of protein is the key step in detection and identification of proteins in all applications of proteomics. The methods currently used for protein extraction from mammalian cells are usually laborious, expensive or insufficiently reliable. Recently, researchers working with mammalian cells conduct protein extraction widely and frequently. This frequent dealing with a large number of samples becomes tedious work. Even though there are many commercial lysis buffers on the market, they are usually expensive. Therefore, in this study, protein extraction lysis buffer was designed to provide highly efficient total protein extraction from cultured mammalian cells. The simple and unique composition of this lysis buffer containing sodium dodecyl sulphate (SDS) dissolves cell membranes and extracts total cellular protein in only 45 minutes. The novel extraction method and lysis buffer composition was formulated using classical optimization or one factor at a time (OFAT) study. In this method, cell pellet from mammalian cells (CHO-K1) were subjected to different lysis buffer treatment procedure and then incubated at room temperature. The SDS-PAGE was performed to confirm the proteins extracted. The method was extended to bacteria (Escherichia coli, BL21-AI), yeast (Pichia pastoris) and repeated for mammalian (CHOK1) for confirmation. It yields complete extraction for bacteria and mammalian cells. As such, this lysis buffer when used for preparing protein extracts from bacteria and mammalian cells offers rapid, reliable and cheap protocol which is applicable to routine laboratory analysis especially for protein estimation and SDS-PAGE.
format	Article
author	Packeer Mohamed, Vasila Hashim, Yumi Zuhanis Has-Yun Amid, Azura Mohd Salleh, Hamzah Yusof, Faridah Mel, Maizirwan
author_facet	Packeer Mohamed, Vasila Hashim, Yumi Zuhanis Has-Yun Amid, Azura Mohd Salleh, Hamzah Yusof, Faridah Mel, Maizirwan
author_sort	Packeer Mohamed, Vasila
title	A rapid method for large sample number extraction of proteins from mammalian cells
title_short	A rapid method for large sample number extraction of proteins from mammalian cells
title_full	A rapid method for large sample number extraction of proteins from mammalian cells
title_fullStr	A rapid method for large sample number extraction of proteins from mammalian cells
title_full_unstemmed	A rapid method for large sample number extraction of proteins from mammalian cells
title_sort	rapid method for large sample number extraction of proteins from mammalian cells
publisher	Tissue Engineering and Regenerative Medicine Society of Malaysia
publishDate	2012
url	http://irep.iium.edu.my/27821/ http://irep.iium.edu.my/27821/ http://irep.iium.edu.my/27821/1/Paper_-_Packeer-Mohamed_et_al_2012_%28RegRes%29.pdf
first_indexed	2023-09-18T20:41:13Z
last_indexed	2023-09-18T20:41:13Z
_version_	1777409378168602624

A rapid method for large sample number extraction of proteins from mammalian cells

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