Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (AB270118)

Flow cytometry overlay histogram showing HeLa cells stained with ab270118 (red line). The cells were fixed with 80 % methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab270118) (1x 10⁶ in 100μL at 0.2μg/mL (1/2500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor 488 ^® (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 4 % formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (AB270118)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Pericentrin with ab270118 at 1/50 dilution.

Positive staining on human tonsil.

The section was incubated with ab270118 for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (AB270118)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling Pericentrin with ab270118 at 1/50 dilution.

Positive staining on human breast.

The section was incubated with ab270118 for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (AB270118)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Pericentrin with ab270118 at 1/50 dilution.

Confocal image showing centriole staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (AB270118)

ab270118 staining Pericentrin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% 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 ab270118 at 1/500 dilution (shown in green) and ab190573, Rabbit monoclonal to alphaTubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).