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Western blot analysis of Raji lysates using FoxP3 antibody |
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Blank control (Black line): Mouse spleen (Black). Primary Antibody (green line): Rabbit Anti-FoxP3 antibody (orb156940), Dilution: 3 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, 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 room temperature. The cells were then incubated in 5% BSA goat serum to block non-specific protein-protein interactions for 15 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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Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-FoxP3 antibody (orb156940), 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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Blank control: MCF7. Primary Antibody (green line): Rabbit Anti-FoxP3 antibody (orb156940), 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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Blank control: Mouse spleen. Primary Antibody (green line): Rabbit Anti-FoxP3/PE Conjugated antibody, Dilution: 0.2 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG-PE. 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. The cells were stained with Primary Antibody for 30 min at room temperature. Acquisition of 20000 events was performed. |
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Sample: Lane 1: PBMC (Mouse) Lysate at 40 ug, Lane 2: Thymus (Mouse) Lysate at 40 ug, Lane 3: Spleen (Mouse) Lysate at 40 ug, Lane 4: Thymus (Rat) Lysate at 40 ug, Primary: Anti-FoxP3 (orb156940) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 43/45 kD, Observed band size: 43/45 kD. |
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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 37C for 20 min, Incubation: Anti-FoxP3 Polyclonal Antibody, Unconjugated (orb156940) 1:500, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: rat spleen, 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-FoxP3 Polyclonal Antibody, Unconjugated (orb156940) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
