Effective separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide by altering conditions of polyacrylamide gel electrophoresis

J P Buyon; S G Slade; E K Chan; E M Tan; R Winchester

doi:10.1016/0022-1759(90)90440-7

Effective separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide by altering conditions of polyacrylamide gel electrophoresis

J Immunol Methods. 1990 May 25;129(2):207-10. doi: 10.1016/0022-1759(90)90440-7.

Authors

J P Buyon¹, S G Slade, E K Chan, E M Tan, R Winchester

Affiliation

¹ Institute of Molecular Immunology, Hospital for Joint Diseases of the New York University Medical Center, New York 10003.

PMID: 2351837
DOI: 10.1016/0022-1759(90)90440-7

Abstract

A method is described for the separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide. These two proteins anomalously co-migrate with the same relative mass under conditions of conventional sodium dodecyl sulfate polyacrylamide gel electrophoresis according to Laemmli using a stock solution of acrylamide: bisacrylamide 30:0.8, ratio = 37.5. A higher ratio of monomer to cross-linker, ratio = 172.4 used in a 15% acrylamide solution, readily separates the two peptide chains. This method facilitates the detection of the 52 kDa SSA/Ro component which otherwise might have been incorrectly assigned as a 48 kDa SSB/La polypeptide.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Autoantigens / analysis*
Autoantigens / isolation & purification
Electrophoresis, Polyacrylamide Gel
Immunoblotting
Molecular Weight
RNA, Small Cytoplasmic*
Ribonucleoproteins*
SS-B Antigen

Substances

Autoantigens
RNA, Small Cytoplasmic
RO60 protein, human
Ribonucleoproteins
SS-A antigen