Blank control: MCF-7. Primary Antibody (green line): Rabbit Anti-MARK2 antibody, Dilution: 1:100, Secondary Antibody: Goat anti-rabbit IgG-AF488, Dilution: 1:1000. Protocol, 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 (human appendix), 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 (MARK2) Monoclonal Antibody, Unconjugated (orb1499351) at 1:50 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney), 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, Incubation with (GEF H1) Monoclonal Antibody, Unconjugated (orb1499351) at 1:100 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney), 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, Incubation with (GEF H1) Monoclonal Antibody, Unconjugated (orb1499351) at 1:100 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
Sample: Lane 1: MCF-7 cell lysate, Lane 2: SK-Br-3 cell lysate, Primary: Anti-MARK2 (orb1499351) at 1:2000 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 88 kD, Observed band size: 77/88 kD.
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