Anti-Pericentrin antibody [EPR21987] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Pericentin with ab220784 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of centrosomes in human pancreas (PMID : 16534625, PMID : 22031837). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

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Multiplex immunohistochemistry - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining PCM1 with ab324615 at a 1 : 200 (2.5 ug/ml) dilution, ab237034 anti-Pericentrin used at 1 : 200 (5.0 ug/ml) dilution.

Panel A : merged staining of anti-PCM1 (green; Opal™520) and anti-Pericentrin (magenta; Opal™570) on human colon.
Panel B : anti-PCM1 staining centrosomes in human colon.
Panel C : ant-Pericentrin staining centrosomes in human colon.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324615 and ab237034 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Immunofluorescent analysis of 100% Methanol-fixed NIH/3T3 (mouse embyro fibroblast cell line) cells labeling Pericentrin with ab220784 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing centrosome staining in NIH/3T3 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Pericentrin with ab220784 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Immunofluorescent analysis of 100% Methanol-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Pericentrin with ab220784 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing centrosome staining in HeLa cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue labeling Pericentin with ab220784 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of centrosomes in human thyroid carcinoma (PMID : 16534625, PMID : 22031837). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Pericentin with ab220784 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining of centrosomes in human mammary epithelial cells (PMID : 16534625, PMID : 22031837). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Pericentrin was immunoprecipitated from 0.35 mg HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab220784 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220784 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HepG2 whole cell lysate 10 μg (input).
Lane 2 : ab220784 IP in HepG2 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220784 in HepG2 whole cell lysate (-).

Blocking and dilution buffer and concentration : 5% NFDM/TBST
Exposure time : 30 seconds

The 220 kDa band represent an isoform of Pericentrin (PMID : 23060948). We used a high sensitivity ECL substrate to develop this blot.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

All lanes:

Immunoprecipitation - Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (<a href='/en-us/products/primary-antibodies/pericentrin-antibody-epr21987-centrosome-marker-ab220784'>ab220784</a>)

Predicted band size: 378 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Pericentrin antibody [EPR21987] - BSA and Azide free (AB237034)

Pericentrin was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate with ab220784 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220784 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10 μg (input).
Lane 2 : ab220784 IP in NIH/3T3 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab220784 in NIH/3T3 whole cell lysate (-).

Blocking and dilution buffer and concentration : 5% NFDM/TBST
Exposure time : 30 seconds

The 220 kDa band represent an isoform of Pericentrin (PMID : 23060948). We use high sensitive ECL substrate.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220784).

All lanes:

Immunoprecipitation - Anti-Pericentrin antibody [EPR21987] - Centrosome Marker (<a href='/en-us/products/primary-antibodies/pericentrin-antibody-epr21987-centrosome-marker-ab220784'>ab220784</a>)

Predicted band size: 378 kDa

Observed band size: 220 kDa

false