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A549 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 (MKLP1) Monoclonal Antibody, Unconjugated (orb704336) 1:50, 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. |
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Blank control: A549. Primary Antibody (green line): Rabbit Anti-MKLP1 antibody (orb704336), dilution: 2 ul/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 90% ice-cold methanol 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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Hela 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 (MKLP1) Monoclonal Antibody, Unconjugated (orb704336) 1:50, 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. |
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Sample: Lane 1: 293 cell lysate, Lane 2: A549 cell lysate, Primary: Anti-MKLP1 (orb704336) at 1:500 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 110 kD, Observed band size: 110 kD. |
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Sample: Lane 1: Rat Lymph node tissue lysates, Lane 2: Rat Cerebrum tissue lysates, Lane 3: Rat Testis tissue lysates, Primary: Anti-MKLP1 (orb704336) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 110 kDa, Observed band size: 100 kDa. |
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Flow cytometry of A549.Primary Antibody (green line): Rabbit Anti-MKLP1 antibody, Dilution:2ul/Test. |
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Western blot:Lane 1:293 Cell,Lane 2:A549 Cell,Primary:Anti-MKLP1 at 1:500 dilution. |