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The pericentrin clone used is 1.7 kb in size and is derived from within residues 100-600 of mouse pericentrin 1. It was expressed as a fusion protein. The corresponding amino acids are present in both pericentrin and kendrin (pericentrin-2) so this antibody is predicted to cross-react with both isoforms.
Our Abpromise guarantee covers the use of ab28144 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Immunocytochemistry/ Immunofluorescence analysis of Y79 cells labeling Pericentrin with ab28144 (red). Cells were fixed for 10 minutes at room temperature in 4% paraformaldehyde. Cells were then permeabilized using 0.2% Triton/TBS and blocked with 5% NGS in 0.1% BSA/TBS-Tween. The cells were then incubated overnight at 4°C with Anti-Pericentrin antibody [mAbcam 28144] - Centrosome Marker (ab28144) at 1/1000 diluted in 0.1% BSA/TBS-Tween containing 1% NGS. Cells were then stained for Hoechst (blue) and alpha-tubulin (green).
ICC/IF image of ab28144 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab28144, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunocytochemistry/ Immunofluorescence analysis of A549 lung cells labeling Pericentrin with ab28144 at 1/100 dilution. Cells were fixed with formaldehyde and permeabilized with Trixton X-100. The cells were blocked with 10% serum for 30 minutes at 20°C, followed by incubation with ab28144 in PBS 10% FCS for 1 hour at 20°C. A polyclonal goat anti-mouse Alexa Fluor® 594 secondary antibody was used at 1/300 dilution.
Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Pericentrin with ab28144 at 1/200 dilution. Cells were fixed with methanol, followed by incubation with Anti-Pericentrin antibody [mAbcam 28144] - Centrosome Marker (ab28144) in PBS for 1 hour at 22°C. A polyclonal goat anti-mouse Cy3® secondary antibody was used at 1/200 dilution. Nuclear counterstained with DAPI.