New Protein Extraction/Solubilization Protocol for Gel-based Proteomics of Rat (Female) Whole Brain and Brain Regions

The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electro...

Full description

Saved in:
Bibliographic Details
Published in:Molecules and cells 2006-08, Vol.22 (1), p.119-125
Main Authors: Hirano, Misato (Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), WEST, Ibaraki, Japan), Rakwal, Randeep (Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), WEST, Ibaraki, Japan), E-mail: rakwal-68@aist.go.jp, Shibato, Junko (Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), WEST, Ibaraki, Japan), Agrawal, Ganesh Kumar (Research Laboratory for Agricultural Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal), Jwa, N.S. (Sejong University, Seoul, Republic of Korea), Iwahashi, Hitoshi (Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), WEST, Ibaraki, Japan), Masuo, Yoshinori (Human Stress Signal Research Center (HSS), National Institute of Advanced Industrial Science and Technology (AIST), WEST, Ibaraki, Japan)
Format: Article
Language:English
Subjects:
Animals
Brain Chemistry
Chemical Fractionation - methods
Electrophoresis, Gel, Two-Dimensional - methods
Female
Gel-based Proteomics
IPG
Lysis Buffer
Nerve Tissue Proteins - isolation & purification
Peptide Mass Fingerprinting
PROTEINAS
PROTEINE
PROTEINS
Proteomics - methods
Rat Brain
Rats
Rats, Wistar
Solubility
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
TCA/Acetone
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins.
ISSN:1016-8478
DOI:10.1016/s1016-8478(23)17399-7