ab94730 staining MLC1 in primary Rat cortical astrocytes by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 4% paraformaldehyde. After 1 hour of incubation with blocking solution (5% BSA in PBS), cells were incubated for 1 hour at room temperature with a NBD phallacidin high-affinity F-actin probe (1/30) to stain actin filaments and visualize the whole cell body, used in combination with ab94730 (1/50) diluted in PBS+0.2% Triton X100.To amplify MLC1 staining, a biotinilated secondary antibody (4.3 µg/ml, Biotin-SP-AffiniPure goat anti-rabbit IgG H+L) followed by incubation with 2 µg/ml streptavidin-TRITC was used.
Western blot - MLC1 antibody (ab94730)
Anti-MLC1 antibody (ab94730) at 1 µg/ml (in 5% skim milk / PBS buffer) + Human fetal spleen lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution