Microheterogeneity of human erythrocyte pyruvate kinase: application of immunological visualization techniques after isoelectric focusing in ultrathin gels.
Immunological methods for protein visualization afford high sensitivity, good specificity and resolution. We used the indirect immunoassay with peroxidase- and 125I-labelled antibodies to investigate the microheterogeneity of pyruvate kinase from human red blood cells and can show an enzyme pattern with more than 10 single bands. Different methods were tested as to their applicability for protein fixation and subsequent immunological visualization in polyacrylamide gels. Besides the Western blot, immunofixation and ethanolic fixation can be recommended as fixation techniques, especially after isoelectric focusing in ultrathin gels.
Megow
D
D
Institute of Biochemistry, Humboldt-University Berlin, German Democratic Republic.
Jacobasch
G
G
D000180:Acrylic Resins; D015153:Blotting, Western; D004912:Erythrocytes / Q000201:enzymology*; D000431:Ethanol; D005404:Fixatives; D006801:Humans; D007118:Immunoassay; D007124:Immunoenzyme Techniques; D007457:Iodine Radioisotopes; D007525:Isoelectric Focusing; D011770:Pyruvate Kinase / Q000097:blood*
D000180:Acrylic Resins; D005404:Fixatives; D007457:Iodine Radioisotopes; C016680:polyacrylamide gels; D000431:Ethanol; D011770:Pyruvate Kinase