The 6/2 (AA3) polyclonal antibody identifying a 37 kD keratinocyte protein reacts also with BM‐600/nicein, the basement membrane component bound by the monoclonal antibody GB3
- 1 February 1992
- journal article
- Published by Wiley in Experimental Dermatology
- Vol. 1 (1) , 52-58
- https://doi.org/10.1111/j.1600-0625.1992.tb00072.x
Abstract
An unexpected finding concerning our previously reported polyclonal antibody raised against an extract from human amnion (pAb 6/2, also termed AA3), and which recognizes an epidermal keratinocyte protein, is presented in this study. Using the immunoblot technique, pAb 6/2 binds to a 37 kD intracellular protein antigen. We have subsequently found that, by radioimmunoprecipitation performed after metabolic labelling with 35S-methionine of cultured keratinocytes, pAb 6/2 recognizes the 600 kD epidermal basement membrane component (termed BM-600/nicein) which was reported to be bound by the monoclonal antibody mAb GB3. Specifically, pAb 6/2 reacts with immunoaffinity chromatography-isolated BM-600/nicein blotted onto nitrocellulose. The data suggest the existence of two immunological reactivities borne by pAb 6/2, each of them being directed against, respectively, the 37 kD (seen in immunoblots) and the 600 kD protein (seen in immunoprecipitations). The data further suggest possible independent expression of these two proteins in cell culture. In comparison with the staining pattern of normal skin, immunofluorescence was previously noted to be impaired (pAb 6/2) or absent (mAb GB3) in lethal junctional epidermolysis bullosa. Thus, we conclude that mAb GB3, rather than pAb 6/2, is a more appropriate probe for the comprehensive biochemical study of this genodermatosis.Keywords
This publication has 13 references indexed in Scilit:
- The new basement membrane antigen recognized by the monoclonal antibody GB3 is a large size glycoprotein: modulation of its expression by retinoic acidBiochimica et Biophysica Acta (BBA) - Biomembranes, 1988
- Monoclonal antibody GB3, a new probe for the study of human basement membranes and hemidesmosomesExperimental Cell Research, 1987
- Identification of a 37 Kilodalton Protein at the Epidermal Basement Membrane by an Antiserum to Human AmnionJournal of Investigative Dermatology, 1986
- GB3 MONOCLONAL ANTIBODY FOR DIAGNOSIS OF JUNCTIONAL EPIDERMOLYSIS BULLOSAThe Lancet, 1986
- Abnormal binding of an anti-amnion antibody to epidermal basement membrane provides a novel diagnostic probe for junctional epidermolysis bullosaBritish Journal of Dermatology, 1985
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Formation of a keratinizing epithelium in culture by a cloned cell line derived from a teratomaCell, 1975
- Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell isCell, 1975
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970