[A method of extraction and separation of wheat gluten].
After removing gliadin and other proteins with 7.5% 2-propanol and 0.3 mol/L NaI, the total glutenin subunits were extracted with two kinds of buffer, one containing 25% 2-propanol, 0.04 mol/L Tris-HCI (pH=8.0), 10% SDS and 2% DDT and the other containing 25% 2-propanol, 0.04 mol/L Tris-HCI (pH=8.0), 10% SDS and 1.4% VP. In SDS-PAGE electrophoresis (4% stacking gel and 13% resolving ge1) system HMW-GS and LMW-GS were clearly separated by the improved method with only a single step. The background of the SDS-PAGE gel was gliadin-free. It was more effective and applicable to separate and distinguish the complex LMW-GS. This method will be used in wheat protein separation, variety identification and improvement.
Ji
Jun
J
Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology, CAS, Shijiazhuang 050021, China, jijun12345@163.com
Liu
Dong-Cheng
DC
Wang
Jing
J
Li
Jun-Ming
JM
Zhang
Ai-Min
AM
D019840:2-Propanol / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005983:Glutens / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D012680:Sensitivity and Specificity; D012974:Sodium Iodide / Q000737:chemistry; D013997:Time Factors; D014908:Triticum / Q000737:chemistry*
D005983:Glutens; D012974:Sodium Iodide; D019840:2-Propanol