Blank control (black line): A549. Primary Antibody (green line): Rabbit Anti-RBM20 antibody (orb1364), Dilution: 1 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488, Dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, 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.
Blank control: A549. Primary Antibody (green line): Rabbit Anti-RBM20 antibody (orb1364), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, Dilution: 3 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 -20C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at 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: heart (mouse) Lysate at 40 ug, Primary: Anti-RBM20 (orb1364) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 75kD, Observed band size: 130 kD.
Sample: K562 (Human) Cell Lysate at 30 ug, A549 (Human) Cell Lysate at 30 ug, Primary: Anti-RBM20 (orb1364) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 134 kD, Observed band size: 130 kD.
Tissue/Cell: mouse embryo 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 37C for 20 min, Incubation: Anti-RBM20 Polyclonal Antibody, Unconjugated (orb1364) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining.
Immunohistochemical staining of mouse embryo tissue using RBM20 antibody.
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