Anti-Pericentrin antibody - Centrosome Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (AB4448)

ab4448 staining Pericentrin in NIH3T3 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab4448 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (AB4448)

IF staining of pericentrin in MCF7 (Human breast adenocarcinoma cell line) cells.

The top panel is an interphase cell showing centrosome staining.

The bottom panel shows a mitotic cell with spindle pole staining.

ab4448 was used at 1/500, but also works at higher dilutions (1/1000-1/2000).

Top panel - 630X magnification; Bottom panel -1000X magnification.

The secondary antibody was Alexa-Fluor^®488 anti-rabbit.

This image is courtesy of Gordon Chan, University of Alberta

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (AB4448)

ab4448 staining Pericentrin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab4448 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (AB4448)

A representative panel of indirect immunofluorescence microscopic images shows normal (regular, n ≤ 2) and aberrant centrosome numbers (n > 2) in interphase cells.

Centrosomes were stained using anti-pericentrin antibody ab4448 (magenta), nuclear DNA is shown in blue (DAPI).

Statistical methods : Kruskal-Wallis test. Mann-Whitney U tests followed by Bonferroni-Holm p-value correction were made as post-hoc tests in order to compare the MDS and sAML patients with the control group.

Ruppenthal et al PLoS One. 2018 Jan 25;13(1):e0191734. doi: 10.1371/journal.pone.0191734. eCollection 2018. Fig 2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (AB4448)

NIH/3T3 (Mouse embryo fibroblast cell line) cells were fixed in 100% methanol for 6 minutes at -20°C, washed 3 times in PBS then incubated with ab4448 (1/2000) for 1 hour at room temperature.

The panel of images shows the nuclei stained with DAPI (blue), ab4448 staining is shown in green. 100x magnification.

This image is courtesy of Roberto Giambruno, Marilena Ciciarello and Patrizia Lavia