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.
This antibody clone is manufactured by Abcam.We can conjugate this antibody to FITC for you (please see ab150205 for details).
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.