The effect of sodium dodecyl sulfate and Pluronic F127 on the electrophoretic separation of protein and polypeptide test mixtures at acid pH.
Using a test mixture consisting of standard proteins (cytochrome c, chymotrypsinogen A, hen egg albumin, bovine serum albumin, aldolase, catalase and ferritin) and synthetic polypeptides (polylysine, polyaspartic, polyglutamic acid and polyproline) it was revealed that using sodium dodecyl sulfate (SDS) as background electrolyte modifier at acid pH (2.5) allows selective separation of highly positively charged polypeptides (polylysine) provided that their relative molecular mass is sufficiently low (3300 Da). The altered elution sequence of standard proteins as compared to a separation done without SDS may help their identification. Addition of Pluronic F127 offers clear-cut separations of standard proteins up to a relative molecular mass of 5 x 10(4) Da and allows to reveal protein/polypeptide microheterogeneity where applicable. None of the systems tested is suitable for the separation of acidic polypeptides and polyproline.
Miksík
Ivan
I
Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. miksik@biomed.cas.cz
Eckhardt
Adam
A
Forgács
Esther
E
Cserháti
Tibor
T
Deyl
Zdenek
Z
D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods; D010455:Peptides / Q000302:isolation & purification*; D010710:Phosphates; D020442:Poloxamer; D011506:Proteins / Q000302:isolation & purification*; D012822:Silicon Dioxide; D012967:Sodium Dodecyl Sulfate; D013501:Surface-Active Agents
D002021:Buffers; D010455:Peptides; D010710:Phosphates; D011506:Proteins; D013501:Surface-Active Agents; D020442:Poloxamer; D012967:Sodium Dodecyl Sulfate; D012822:Silicon Dioxide