25 ug total protein per lane of various lysates (see on figure) probed with GFAP polyclonal antibody, unconjugated (bs-42001R) at 1:20000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

4% Paraformaldehyde-fixed U-251 MG (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (GFAP) polyclonal Antibody, unconjugated (bs-42001R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

The U-251 MG (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-GFAP antibody (bs-42001R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

Measured by its binding ability in a indirect ELISA. Immobilized Human GFAP protein, His Tag (Cat. bs-42001P) at 2 μg/mL (100 μL/well) can bind  Rabbit Anti-Human GFAP Antibody, the EC50 is 540.1 ng/mL.