Xenbase Image ID: 151301

Fig. 2. Western immunoblot analysis of A6 cells and embryo residues. Whole cell and detergent-insoluble residues of A6 cells were separated by SDS-polyacrylamide gel electrophoresis and either stained with Coomassie brilliant blue (A) (we, whole cell; cr, cell residue) or were electrophoretically transferred to nitrocellulose paper and probed with the monoclonal antibody RV202 (B) or 14h7 (C). RV202 labelled a single band of 55xlO in both whole cell (we) and cell residue (cr) lanes. Occasionally a band below the 55 x 103 polypeptide was labelled (asterisk), this is presumably a proteolytic degradation product of the 55 x 103 polypeptide. Typically 14h7 appears to react weakly with a 55x10 polypeptide in whole cell, and with polypeptides of 33 55xlO (large arrow) and 57xlO (small arrow) in cell residue (C). To illustrate that RV202 and 14h7 react with 3 the same 55xlO polypeptide, a single lane of A6 cell residue was cut down the middle (D) and probed with 14h7 3 (left side) or RV202 (right side); the labelled 55xlO polypeptides align perfectly. To define the specificity of 14h7 and RV202 in embryos, insoluble residues from stage- 35 tadpoles were separated by SDS-polyacrylamide gel electrophoresis and either stained with Coomassie brilliant blue (E) or transferred to nitrocellulose and probed with either RV202 (F) or 14h7 (G). RV202 reacted with poly- 3 peptide of 55xlO , 14h7 reacted with this same polypeptide (large arrow), and very weakly with a polypeptide of 3 57xlO . A minor band that presumably represents a proteolytic degradation product of the 55 x 103 polypeptide was also labelled by both antibodies (asterisk in F and G). Dash marks on the left side of A and E mark the position 3 of the molecular weight markers phosphorylase (97xlO ), 33 bovine serum albumin (66X10 ), ovalbumin (45xlO ) and 3 carbonic anhydrase (29x10 3) Image published in: Dent JA et al. (1989) Copyright © 1989. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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