Western blot using the Protein-A Purified anti-bovine VEGF-A antibody shows detection of recombinant bovine VEGF-A at 17-19.2 kDa (arrowhead). Approximately 2 µg of recombinant protein was loaded per lane onto a 4-20% gradient gel followed by transfer to PVDF membrane. The membrane was blocked using 3% BSA diluted 1:10. The primary antibody was used at a 1:333 dilution and was incubated with the blot for 2h at room temperature. The membrane was washed and reacted with a 1:10,000 dilution of Conjugated Affinity Purified Goat-anti-Rabbit IgG [H&L] MX10. Molecular weight estimation was made by comparison to prestained MW markers. Other detection systems will yield similar results.