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Blank control (blue line): Mouse spleen cells (blue). Primary Antibody (green line): Rabbit Anti-Cyclin E1 antibody (orb10499), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 70% ethanol (overninght at 4C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed. |
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Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-Cyclin E1 antibody (orb10499), 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 -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. |
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Cell: NIH/3T3, Concentration: 1:100, Host/Isotype: Rabbit/IgG, Flow cytometric analysis of primary antibody (Cat: orb10499) on NIH/3T3 (green) compared with Rabbit IgG isotype control in the absence of primary antibody (blue) followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG (H+L) secondary antibody. |
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Sample: Lovo (Human) Cell Lysate at 30 ug, Thymus (Mouse) Lysate at 40 ug, U2os (Human) Cell Lysate at 30 ug, K562 (Human) Cell Lysate at 30 ug, Primary: Anti-Cyclin E1 (orb10499) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 45 kD, Observed band size: 50 kD. |
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Tissue/Cell: human laryngocarcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), 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-Cyclin-E Polyclonal Antibody, Unconjugated (orb10499) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: rat testis tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37C for 20 min, Incubation: Anti-Cyclin E Polyclonal Antibody, Unconjugated (orb10499) 1:200, overnight at 4C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (orb868589) used at 1:200 dilution for 40 minutes at 37C. |
