Total protein extraction with TCA-acetone.
We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneous precipitation and denaturation with TCA and 2ME in cold acetone. We also describe protein solubilization prior to IEF, either in classical rod gels or in IPGs, using two different solutions. The procedure is easy to carry out. The major caveats are (1) keep samples at low temperature during extraction, and then (2) manage protein samples at about 22 to 25 degrees C to avoid urea precipitation.
Méchin
Valérie
V
UMR de Chimie Biologique, Thiverval Grignon, France.
Damerval
Catherine
C
Zivy
Michel
M
D000096:Acetone / Q000737:chemistry*; D011232:Chemical Precipitation; D003902:Detergents / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D013696:Temperature; D014238:Trichloroacetic Acid / Q000737:chemistry*
D003902:Detergents; D010940:Plant Proteins; D000096:Acetone; D014238:Trichloroacetic Acid