VALIDATION IMAGES
25 ug total protein per lane of various lysates (see on figure) probed with TFRC monoclonal antibody, unconjugated (bsm-43165M) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
The Hela (H) cells 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): Mouse Anti-TFRC antibody (bsm-41365M): 1 μg/10^6 cells; Isotype Control (orange): Mouse IgG (bs-0296P). Secondary Antibody (white blue): Goat anti-Mouse IgG-FITC (bs-40296G-FITC): 1 μg/test.Blank control (black): PBS. Acquisition of 20,000 events was performed.
HeLa cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (TFRC) monoclonal Antibody, Unconjugated (bsm-43165M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (TFRC) monoclonal Antibody, unconjugated (bsm-43165M) 1:200, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Mouse IgG antibody (green, bs-60296G-BF488) at 37°C for 90 min, then stained with Phalloidin/BF555 for 30 min at room temperature; DAPI (blue, C02-04002) was used to stain the cell nuclei.