ASCL1 Rabbit Polyclonal Antibody

A549 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (MASH1) polyclonal Antibody, Unconjugated (orb10143) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-MASH1 antibody (orb10143), Dilution: 2 ug/Test, Secondary Antibody: Goat anti-rabbit 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 -20℃. 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.

Blank control: Mouse brain. Primary Antibody (green line): Rabbit Anti-MASH1 antibody (orb10143), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/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 -20℃. 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.

Sample: Brain (Mouse) Lysate at 40 ug Primary: Anti-ASCL1 (orb10143) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 26 kD Observed band size: 26 kD.

Tissue/cell: human glioma tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37℃ for 20 min, Incubation: Anti-MASH1 Polyclonal Antibody, Unconjugated (orb10143) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/cell: mouse embryos tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37℃ for 20 min, Incubation: Anti-MASH1 Polyclonal Antibody, Unconjugated (orb10143) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37℃ for 20 min, Incubation: Anti-MASH1 Polyclonal Antibody, Unconjugated (orb10143) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/cell: Rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37℃ for 20 min, Incubation: Anti-MASH1 Polyclonal Antibody, Unconjugated (orb10143) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.