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Blank control: U937 (blue). Primary Antibody: Rabbit Anti-Integrin Alpha V + Beta 3 (CD51+CD61) antibody (orb10927), Dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde (10 min). Primary antibody (orb10927, 1 µg/1x10 6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20000 events was performed. |
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Cell: MCF-7, Concentration: 1:100, Host/Isotype: Rabbit/IgG, Flow cytometric analysis of Rabbit IgG isotype control (Cat: orb10927) on MCF-7 (green) compared with 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: HUVEC (human) cell Lysate at 40 ug, Primary: Anti-Integrin Alpha V + Beta 3 (CD51+CD61) (orb10927) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 116 kD, Observed band size: 120 kD. |
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Tissue/Cell: Human laryngeal tissue, 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-Integrin Alpha V + Beta 3 (CD51 + CD61) Polyclonal Antibody, Unconjugated (orb10927) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: Human lung cancer tissue, 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-Integrin Alpha V + Beta 3 (CD51 + CD61) Polyclonal Antibody, Unconjugated (orb10927) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
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Tissue/Cell: rat brain tissue, 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-Integrin Alpha V + Beta 3 (CD51+CD61) Polyclonal Antibody, Unconjugated (orb10927) 1:200, overnight at 4C, followed by conjugation to the secondary antibody and DAB staining. |
