Anti-UHRF1 antibody [EPR18803-11]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling UHRF1 with ab213223 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^®488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Goat anti-Mouse IgG (Alexa Fluor^® 594)) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab213223 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling UHRF1 with ab213223 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on Human tonsil germinal center is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling UHRF1 with ab213223 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^®488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Goat anti-Mouse IgG (Alexa Fluor^® 594)) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab213223 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling UHRF1with ab213223 at 1/70 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling UHRF1 with ab213223 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on Human colon cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

UHRF1 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab213223 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab213223 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : HeLa whole cell lysate, 10μg (Input).

Lane 2 : ab213223 IP in HeLa whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab213223 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-UHRF1 antibody [EPR18803-11] (ab213223)

Predicted band size: 89 kDa

false

• WB

Lab

Western blot - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. UHRF1 has two isoforms. UHRF1 undergo degradation under normal conditions as well as in response to DNA damage. (PMID : 23297342, PMID : 36593255)

All lanes:

Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human thymus tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 89 kDa

false

Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 and 2 : 5 seconds; Lane 3 : 30 seconds.

All lanes:

Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 89 kDa

Observed band size: 97 kDa

false

• WB

Lab

Western blot - Anti-UHRF1 antibody [EPR18803-11] (AB213223)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as GAPDH loading control. UHRF1 has two isoforms. UHRF1 undergo degradation under normal conditions as well as in response to DNA damage. (PMID : 23297342, PMID : 36593255)

All lanes:

Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

Human colon tissue lysate at 20 µg

Lane 5:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

U-937 (Human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 89 kDa

false

Exposure time: 20s