Anti-Bovine VEGF-A Antibody - Western Blot. Western blot of Protein-A Purified anti-bovine VEGF-A antibody shows detection of recombinant bovine VEGF-A at 17-19.2 kD (arrowhead). Approximately 2 ug 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 (BSA-30) 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 1:10000 dilution of IRDye800 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.