25 ug total protein per lane of various lysates probed with beta-Actin monoclonal antibody, unconjugated (orb499643) at 1:1000 dilution and 4C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Blank control: Hela. Primary Antibody (green line): Mouse Anti-beta-Actin antibody (orb499643), dilution: 1 ug/Test, Secondary Antibody: Goat anti-Mouse IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Paraformaldehyde-fixed, paraffin embedded (mouse colon), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (orb499643) at 1:200 overnight at 4C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse stomach tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (beta-Actin) Monoclonal Antibody, Unconjugated (ascites of orb499643) at 1:2000 overnight at 4C, followed by a conjugated secondary for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (orb499643) at 1:200 overnight at 4C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
Immunohistochemical staining of mouse colon using Beta-Actin antibody.
Immunohistochemical staining of rat colon using Beta-Actin antibody.
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