Rat lung, cut into 1-2 mm pieces, was cultured for 4 days in 25-30 mL of RPMI supplemented with 10% fetal bovine serum and stimulated with 2.5 ng/mL LPS. The cell culture supernatant was removed, assayed for rat VEGF/VEGFA, and measured 3292.9 pg/mL. Rat spleen, cut into 1-2 mm pieces, was cultured for 3-4 days in 25-30 mL of RPMI supplemented with 10% fetal bovine serum and stimulated with 10 μg/mL ConA. The cell culture supernatant was removed, assayed for rat VEGF, and measured 433.1 pg/mL.

Serum of eight rats was measured. The VEGF/VEGFA concentration of one sample was determined to be 34.3 pg/mL, and the range of VEGF 164 concentration was 0 - 34.3 pg/mL.

Sandwich ELISA standard curve of MP00598-3, Rat VEGF/VEGFA Recombinant Matched Antibody Pair - PBS only. 83610-1-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Eg0941. 83610-2-PBS was HRP conjugated as the detection antibody. Range: 31.25-2000 pg/mL.

Biolayer interferometry (BLl) kinetic assays of 84610-2-PBS against Rat VEGFA were performed. The affinity constant is below 1 pM.