Rehydratable agarose gels: application to isoelectric focusing in 9 molar urea.
A method is described for the preparation of rehydratable agarose gels, with specific application to the direct incorporation of 9 M urea and carrier ampholytes into rehydratable agarose gels for use in isoelectric focusing. After drying the agarose gel containing an uncharged linear polyacrylamide, one gel volume of a 9 M urea-carrier ampholyte solution is absorbed directly into the gel in 60 min, eliminating equilibration or dialysis of the gel in larger volumes of the 9 M urea-carrier ampholyte solution. Proteins with a molecular mass of 970,000 Da can be separated by isoelectric focusing in these rehydratable gels. The focused proteins can then be quantitatively transferred to nitrocellulose in less than 10 min, and any immunostaining procedure can be used to probe the blotted proteins. These agarose gels are easy to make, they rehydrate rapidly and they can be used in applications other than isoelectric focusing.
Hoffman
W L
WL
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas.
Jump
A A
AA
Kelly
P J
PJ
Elanogovan
N
N
D000180:Acrylic Resins; D006801:Humans; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D007525:Isoelectric Focusing / Q000379:methods*; D012685:Sepharose; D012996:Solutions; D014508:Urea; D000099166:Polyacrylamides
D000180:Acrylic Resins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D012685:Sepharose; D012996:Solutions; D014508:Urea; D000099166:Polyacrylamides