Fig. 5: Antibody validation by western blotting. | Communications Biology

Fig. 5: Antibody validation by western blotting.

From: Epitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation

Fig. 5

a Recombinant hANKRD1 (A; 10 ng per well) and total protein (50 µg per well) isolated from whole-cell lysates of adult rat heart tissue (Sham, S), myocardial infarcted Day 2 tissue (infarcted region, I; border region, B; remote region, R) and neonatal rat cardiomyocytes (N), were separated by SDS-PAGE, transblotted onto PVDF membranes and probed with antibody indicated below each panel. M, All-Blue Precision Protein Standards (BioRad) showing only the 50 and 37 kDa marker bands. b Western blot analysis of lane 1. purified recombinant hANKRD1 (Origene Technologies, 4 ng/well); lane 2. lysate from pCMV-Myc transfected H9c2 cells; lane 3. lysate from pCMV-Myc/hANKRD1-transfected H9c2 cells; lane 4. lysate from E. coli cells expressing hANKRD1, probed with antibodies indicated at the bottom of each panel. Total protein loading for H9c2 and E. coli lysates were 20 µg per well.

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