Two-dimensional immobilized metal affinity electrophoresis for capturing a phosphoprotein.
A two-dimensional immobilized metal affinity electrophoresis method is described here. In this method, ferric ions are immobilized in the second-dimensional polyacrylamide gel to extract the phosphoprotein β-casein from a mixture containing proteins with a broad range of pI and MW. Native 7.5-15% gradient tris-glycine gel with SDS tris-glycine gel running buffer are used so that proteins can be separated according to their molecular mass in the second dimension.
Protein Research Laboratory, Research Resources Center, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
Shalini
S
Gupta
G D
GD
Lasanthi
P
P
Jayathilaka
Jin-Sheng
JS
Huang
Bao-Shiang
BS
Lee
D002364:Caseins / Q000302:isolation & purification / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D010750:Phosphoproteins / Q000302:isolation & purification*
D002364:Caseins; D010750:Phosphoproteins