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Counterimmunoblotting: detection of non-denatured or denatured antigens in antibody-containing agarose gels following polyacrylamide gel electrophoresis.
- Source :
-
Journal of immunological methods [J Immunol Methods] 1984 May 25; Vol. 70 (2), pp. 211-9. - Publication Year :
- 1984
-
Abstract
- Utilizing the principles of counterimmunoelectrophoresis, a technique was devised to immunologically identify non-denatured protein antigens resolved by polyacrylamide gel electrophoresis. Proteins were electrophoretically transferred from polyacrylamide gel to antibody-containing agarose gel in a commercially available blotting apparatus. Detection of the antigen/antibody complex by Coomassie blue staining (in the ng range for unlabeled antigen) or autoradiography (in the pg range for radiolabeled antigen) established the sensitivity and specificity of this method. Similar sensitivity could be obtained following conventional sodium dodecyl sulfate polyacrylamide gel electrophoresis after partial removal of the detergent and possible renaturation of proteins. Placement of a nitrocellulose sheet on the anodal side of the agarose gel produced a 'negative replica' (proteins/polypeptides not reacting with specific antibodies in the agarose gel). Counterimmunoblotting provides an additional method for molecular weight determination of protein antigens with sensitivity and specificity comparable to established nitrocellulose blotting (unlabeled proteins) or immunoprecipitation (labeled proteins) procedures.
- Subjects :
- Antigens immunology
Collodion
Electrophoresis, Polyacrylamide Gel
Gels
Humans
Ovalbumin immunology
Paper
Protein Denaturation
Sarcoma Virus, Woolly Monkey immunology
Serum Albumin immunology
Antigen-Antibody Complex
Antigens analysis
Counterimmunoelectrophoresis methods
Immunoelectrophoresis methods
Sepharose analysis
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 70
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 6327826
- Full Text :
- https://doi.org/10.1016/0022-1759(84)90186-8