Low igg bovine serum albumin bsa

Serum was collected from immunized mice by cheek bleeding and analyzed for anti-poxvirus IgG using a standard ELISA protocol. Briefly, serum was serially diluted (1:500, 1:1,000, 1:2,000, 1:4,000, 1:8,000, and 1:16,000) in 1x PBS supplemented with 1% low-IgG bovine serum albumin (BSA; Gemini Bio Products). Each dilution (plated in triplicate) was then incubated in high binding EIA/RIA plates (Corning) that had been pre-coated with 12,500 PFU VACV (Western Reserve strain) per well. Plates were washed with PBS+0.01% Tween (PBST) and incubated with peroxidase labeled anti-mouse IgG (H+L) (Vector Laboratories) at 1:200 dilution in PBS/BSA as above. Plates were developed using ABST Peroxidase Substrate (KPL) and read at 405 nm.

Serum was collected from primed mice by retro-orbital eye bleeding and analyzed for IgG titer by ELISA. Briefly, serum was serially diluted in PBS supplemented with 1% low-IgG bovine serum albumin (BSA; Gemini Bio-Products), ranging from 1:500 to 1:32,000, and incubated in high binding EIA/RIA plates (Corning) pre-coated with 2 HAU of PR8. Plates were then washed with PBS + 0.01% Tween (PBST) and incubated with peroxidase labeled anti-mouse IgG (H+L) (Vector Laboratories, catalog no. PI-2000) at 1:1500 dilution in PBS/BSA (1%). Plates were developed using ABST Peroxidase Substrate (KPL) and read at detection wavelength of 405 nm.