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Blank control (blue line): Hela (blue). Primary Antibody (green line): Rabbit Anti-MMP2 antibody (orb11061), 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 80% methanol (5 min at -20C) 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 (blue line): Hela (fixed with 80% methanol (5 min at -20C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). Primary Antibody (green line): Rabbit Anti-MMP2 antibody (orb11061), 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. |
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Paraformaldehyde-fixed, paraffin embedded (Mouse brain), 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 (MMP2) Polyclonal Antibody, Unconjugated (orb11061) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining. |
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Paraformaldehyde-fixed, paraffin embedded (Rat brain), 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 (MMP2) Polyclonal Antibody, Unconjugated (orb11061) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining. |
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Paraformaldehyde-fixed, paraffin embedded Human Glioma, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with MMP2 Polyclonal Antibody, Unconjugated (orb11061) at 1:400 overnight at 4C, followed by conjugation to the SP Kit (Rabbit) and DAB staining. |
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Paraformaldehyde-fixed, paraffin embedded Mouse Placenta, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with MMP2 Polyclonal Antibody, Unconjugated (orb11061) at 1:400 overnight at 4C, followed by conjugation to the SP Kit (Rabbit) and DAB staining. |
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Sample: A431 (Human) Cell Lysate at 30 ug, Mcf-7 (Human) Cell Lysate at 30 ug, Primary: Anti-MMP-2 (orb11061) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62 kD, Observed band size: 62 kD. |
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Sample: SY5Y (Human) Cell Lysate at 30 ug, U2os (Human) Cell Lysate at 30 ug, U251 (Human) Cell Lysate at 30 ug, Siha (Human) Cell Lysate at 30 ug, Primary: Anti-MMP-2 (orb11061) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62 kD, Observed band size: 60 kD. |
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Tissue/Cell: U-87MG cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37C for 20 min, Antibody incubation with (MMP2) polyclonal Antibody, Unconjugated 1:100, 90 minutes at 37C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37C for 90 minutes, DAPI (blue) was used to stain the cell nuclei. |