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Blank control: A549. Primary Antibody (green line): Rabbit Anti-Cathepsin L antibody (orb10252), Dilution: 1 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 0.1% PBST for 20 min at room temperature. 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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Paraformaldehyde-fixed, paraffin embedded (Human 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, Antibody incubation with (Cathepsin L) Polyclonal Antibody, Unconjugated (orb10252) at 1:200 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining. |
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Protein: line1, rat brain lysates, 30 ug, line2, rat liver lysates, 30 ug, Primary: Anti-Cathepsin L (orb10252) at 1:200, Secondary: HRP conjugated Goat-Anti-Rabbit IgG (orb572747) at 1: 3000, ECL excitated the fluorescence, Predicted band size: 37kD, Observed band size: 19kD, 37kD. |
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Sample: A549 Cell (Human) Lysate at 30 ug, K562 Cell (Human) Lysate at 30 ug, Primary: Anti-Cathepsin L (orb10252) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 19/30/37 kD, Observed band size: 40 kD. |
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Sample: Lane 1: Human HUVEC cell Lysates, Lane 2: Human K562 cell Lysates, Lane 3: Human MCF-7 cell Lysates, Primary: Anti-Cathepsin L (orb10252) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 19/30/37kDa, Observed band size: 40kDa. |
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Tissue/cell: human cervical carcinoma, 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°C for 20 min, Incubation: Anti-Cathepsin Polyclonal Antibody, Unconjugated (orb10252) 1:500, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |