Name: Anti-UHRF1 antibody [EPR18803-11]
Brand: RabMAb
SKU: ab213223
Rating: 5 (2 reviews)

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Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18803-11] to UHRF1
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
Reacts with: Human

Carrier Free

Product name

Anti-UHRF1 antibody [EPR18803-11]
See all UHRF1 primary antibodies
Description

Rabbit monoclonal [EPR18803-11] to UHRF1
Host species

Rabbit
Specificity

UHRF1 is aboundant in tissues like thymus, bone marrow. For other tissues with low expression level, please upload higher amount of tissue lysate or use lower antibody dilution.
Tested applications

Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human colon and thymus lysates; HeLa, Jurkat, U-937 and HepG2. IHC-P: Human tonsil and colon cancer tissues. ICC/IF: Jurkat and HeLa cells. Flow Cyt (intra): Jurkat cells. IP: HeLa whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18803-11
Isotype

IgG
Research areas

Alternative Versions
- Anti-UHRF1 antibody [EPR18803-11] - BSA and Azide free (ab240388)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human UHRF1 protein (ab198770)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab213223 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(1)	1/1000. Detects a band of approximately 97 kDa (predicted molecular weight: 89 kDa).
IHC-P	(1)	1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommended for human only
ICC/IF		1/500. Recommended for human only
IP		1/30.
Flow Cyt (Intra)		1/70.

Notes
WB 1/1000. Detects a band of approximately 97 kDa (predicted molecular weight: 89 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommended for human only
ICC/IF 1/500. Recommended for human only
IP 1/30.
Flow Cyt (Intra) 1/70.

Function

Putative E3 ubiquitin-protein ligase. May participate in methylation-dependent transcriptional regulation. Binds to inverted 5'-CCAAT-3' box 2 in the TOP2A promoter, and activates TOP2A expression. Important for G1/S transition. May be involved in DNA repair and chromosomal stability.
Tissue specificity

Expressed in thymus, bone marrow, testis, lung and heart. Overexpressed in breast cancer.
Pathway

Protein modification; protein ubiquitination.
Sequence similarities

Contains 1 PHD-type zinc finger.
Contains 2 RING-type zinc fingers.
Contains 1 ubiquitin-like domain.
Contains 1 YDG domain.
Developmental stage

Expressed in fetal thymus, liver and kidney.
Domain

The RING finger is required for ubiquitin ligase activity.
The YDG domain mediates the interaction with histone H3.
Post-translational
modifications

Phosphorylated on serine residues. Phosphorylation may enhance DNA-binding activity.
Ubiquitinated; which leads to proteasomal degradation. Polyubiquitination may be stimulated by DNA damage.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q96T88 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 29128 Human
- Omim: 607990 Human
- SwissProt: Q96T88 Human
- Unigene: 108106 Human
Alternative names
- Ac2-121 antibody
- AL022808 antibody
- E3 ubiquitin-protein ligase UHRF1 antibody
- EC 6.3.2.- antibody
- FLJ21925 antibody
- hNP95 antibody
- hUHRF1 antibody
- HuNp95 antibody
- ICBP90 antibody
- Inverted CCAAT box binding protein of 90 kDa antibody
- Inverted CCAAT box binding protein, 90-kD antibody
- Inverted CCAAT box-binding protein of 90 kDa antibody
- Liver regeneration-related protein LRRG126 antibody
- MGC138707 antibody
- NP95 antibody
- Nuclear phosphoprotein 95 antibody
- Nuclear phosphoprotein, 95-KD antibody
- Nuclear protein 95 antibody
- Nuclear zinc finger protein Np95 antibody
- RING finger protein 106 antibody
- RNF106 antibody
- TDRD22 antibody
- Transcription factor ICBP90 antibody
- Ubiquitin like containing PHD and RING finger domains protein 1 antibody
- Ubiquitin like PHD and RING finger domain containing protein 1 antibody
- Ubiquitin-like PHD and RING finger domain-containing protein 1 antibody
- Ubiquitin-like protein containing PHD and RING finger domains 1 antibody
- Ubiquitin-like with PHD and ring finger domains 1 antibody
- Ubiquitin-like, containing PHD and RING finger domains, 1 antibody
- Ubiquitin-like-containing PHD and RING finger domains protein 1 antibody
- UHRF1 antibody
- UHRF1_HUMAN antibody
see all

Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223)

All lanes : Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/1000 dilution

Lane 1 : Human colon tissue lysate
Lane 2 : Human thymus tissue lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 89 kDa
Observed band size: 30-70,90-100 kDa why is the actual band size different from the predicted?

Exposure time: 60 seconds

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)
Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223)

All lanes : Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/1000 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human kidney tissue lysate
Lane 3 : Human spleen tissue lysate
Lane 4 : Human colon tissue lysate
Lane 5 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 6 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 7 : U-937 (Human histiocytic lymphoma monocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 89 kDa
Observed band size: 30-70,90-100 kDa why is the actual band size different from the predicted?

Exposure time: 60 seconds

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)
Western blot - Anti-UHRF1 antibody [EPR18803-11] (ab213223)

All lanes : Anti-UHRF1 antibody [EPR18803-11] (ab213223) at 1/5000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 89 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1 and 2:5 seconds; Lane 3: 30 seconds.
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.
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.
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.
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 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.
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.
Anti-UHRF1 antibody [EPR18803-11] (ab213223)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab213223? Please let us know so that we can cite the reference in this datasheet.

ab213223 has been referenced in 2 publications.

Li Q et al. UHRF1 Knockdown Attenuates Cell Growth, Migration, and Invasion in Cutaneous Squamous Cell Carcinoma. Cancer Invest 39:84-97 (2021). PubMed: 33058714
Chen X et al. Overexpression of UHRF1 promoted the proliferation of vascular smooth cells via the regulation of Geminin protein levels. Biosci Rep 39:N/A (2019). PubMed: 30710064

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1-2 of 2 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (Human ES Cells)

Gel Running Conditions

Reduced Denaturing (4-14% Nupage Bis-Tris)

Loading amount

308000 cells

Specification

Human ES Cells

Blocking step

Li-Cor Odyssey PBS Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Timothy Hunt

Verified customer

Submitted Sep 12 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Cell (Gonad)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9

Permeabilization

Yes - 0.05% triton X in PBS

Specification

Gonad

Blocking step

No blocking step used for 30 hour(s) and 0 minute(s) · Concentration: 10 · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Timothy Hunt

Verified customer

Submitted Aug 23 2017

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Sequence similarities

Developmental stage

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Western blot abreview for Anti-UHRF1 antibody [EPR18803-11]

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

Post-translational
modifications