alpha smooth muscle Actin Recombinant Antibody

25 ug total protein per lane of various lysates (see on figure) probed with alpha smooth muscle Actin monoclonal antibody, unconjugated (bsm-33187R) at 1:20000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

4% Paraformaldehyde-fixed NIH\/3T3 treated with 5 ng\/mL TGF-\u03b21 for 72h (M) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (alpha smooth muscle Actin) monoclonal Antibody, unconjugated (bsm-33187R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C 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 NIH\/3T3 treated with 5 ng\/mL TGF-\u03b21 for 72h (M) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-alpha smooth muscle Actin antibody (bsm-33187R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

4% Paraformaldehyde-fixed WI-38 (treated with 5 ng\/mL TGF-\u03b21 for 72h) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (alpha smooth muscle Actin) monoclonal Antibody, unconjugated (bsm-33187R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C 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 WI-38 ( treated with 5 ng\/mL TGF-\u03b21 for 72h ) (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\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-alpha smooth muscle Actin antibody (bsm-33187R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.