Recombinant Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Product name

Anti-HUWE1/Mule antibody [EPR24361-13]
See all HUWE1/Mule primary antibodies
Description

Rabbit monoclonal [EPR24361-13] to HUWE1/Mule
Host species

Rabbit
Tested applications

Suitable for: IHC-P, ICC, Flow Cyt, WBmore details
Unsuitable for: IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, MOLT-4, 293T, NIH/3T3 and PC-12 whole cell lysate. Mouse heart tissue lysate. IHC-P: Human breast carcinoma and Mouse cardiac muscle and Rat kidney tissue. ICC: HeLa, F9 and MOLT-4 cells. Flow Cyt: MOLT-4 and F9 cells.
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. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24361-13
Isotype

IgG
Research areas

Alternative Versions
- Anti-HUWE1/Mule antibody [EPR24361-13] - BSA and Azide free (ab282738)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab271032 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
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC		1/50.
Flow Cyt		1/50.
WB		1/1000. Predicted molecular weight: 481 kDa.

Notes
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC 1/50.
Flow Cyt 1/50.
WB 1/1000. Predicted molecular weight: 481 kDa.

Application notes

Is unsuitable for IP.

Function

E3 ubiquitin-protein ligase which mediates ubiquitination and subsequent proteasomal degradation of target proteins. Regulates apoptosis by catalyzing the polyubiquitination and degradation of MCL1. Mediates monoubiquitination of DNA polymerase beta (POLB) at 'Lys-41', 'Lys-61' and 'Lys-81', thereby playing a role in base-excision repair. Also ubiquitinates the p53/TP53 tumor suppressor and core histones including H1, H2A, H2B, H3 and H4. Binds to an upstream initiator-like sequence in the preprodynorphin gene. Regulates neural differentiation and proliferation by catalyzing the polyubiquitination and degradation of MYCN. May regulate abundance of CDC6 after DNA damage by polyubiquitinating and targeting CDC6 to degradation.
Tissue specificity

Weakly expressed in heart, brain and placenta but not in other tissues. Expressed in a number of cell lines, predominantly in those from colorectal carcinomas.
Pathway

Protein modification; protein ubiquitination.
Involvement in disease

Defects in HUWE1 are the cause of mental retardation syndromic X-linked Turner type (MRXST) [MIM:300706]; also known as mental retardation and macrocephaly syndrome. MRXST shows clinical variability. Associated phenotypes include macrocephaly and variable contractures.
A chromosomal microduplication involving HUWE1 and HSD17B10 is the cause of mental retardation X-linked type 17 (MRX17) [MIM:300705]; also known as mental retardation X-linked type 31 (MRX31). Mental retardation is characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. In contrast to syndromic or specific X-linked mental retardation which also present with associated physical, neurological and/or psychiatric manifestations, intellectual deficiency is the only primary symptom of non-syndromic X-linked mental retardation.
Sequence similarities

Belongs to the TOM1/PTR1 family.
Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
Contains 1 UBA domain.
Contains 1 UIM (ubiquitin-interacting motif) repeat.
Contains 1 WWE domain.
Domain

The HECT domain mediates inhibition of the transcriptional activity of p53.
Post-translational
modifications

Phosphorylated on tyrosine; phosphorylation is probably required for its ability to inhibit TP53 transactivation.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localization

Cytoplasm. Nucleus. Mainly expressed in the cytoplasm of most tissues, except in the nucleus of spermatogonia, primary spermatocytes and neuronal cells (By similarity). Predominantly cytosolic or perinuclear in some colorectal carcinoma cells.
Target information above from: UniProt accession Q7Z6Z7 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10075 Human
- Entrez Gene: 59026 Mouse
- SwissProt: Q7Z6Z7 Human
- SwissProt: Q5BMM7 Mouse
- SwissProt: Q7TMY8 Mouse
- SwissProt: P51593 Rat
- Unigene: 136905 Human
- Unigene: 27372 Mouse
- Unigene: 12130 Rat
see all
Alternative names
- ARF binding protein 1 antibody
- ARF BP1 antibody
- ARF-binding protein 1 antibody
- ARF-BP1 antibody
- ARFBP1 antibody
- BJ-HCC-24 tumor antigen antibody
- E3 ubiquitin protein ligase HUWE1 antibody
- E3 ubiquitin-protein ligase HUWE1 antibody
- HECT antibody
- HECT domain protein LASU1 antibody
- HECT UBA and WWE domain containing protein 1 antibody
- HectH9 antibody
- Homologous to E6AP carboxyl terminus homologous protein 9 antibody
- Huwe1 antibody
- HUWE1_HUMAN antibody
- Ib772 antibody
- Large structure of UREB1 antibody
- LASU1 antibody
- Mcl 1 ubiquitin ligase E3 antibody
- Mcl-1 ubiquitin ligase E3 antibody
- Mule antibody
- UBA and WWE domain-containing protein 1 antibody
- Upstream regulatory element-binding protein 1 antibody
- URE-B1 antibody
- URE-binding protein 1 antibody
- UREB1 antibody
see all

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

All lanes : Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 481 kDa
Observed band size: 482 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST. The expression pattern & observed MW are consistent with what has been described in the literature (PMID: 25883150, 28938460).

Low expression: MOLT-4 (PMID: 15567145).

Exposure time: 5.5 seconds
Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

All lanes : Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 481 kDa
Observed band size: 482 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBSTLysates were made freshly and used in WB test immediately to minimize protein degradation.

The expression pattern & observed MW are consistent with what has been described in the literature (PMID: 28938460).

Exposure time: Lane1: 3.25 seconds. Lane2-3: 5.5 seconds
Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution + Mouse heart tissue lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 481 kDa
Observed band size: 482 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST. The expression pattern & observed MW are consistent with what has been described in the literature (PMID: 28938460).

Exposure time: 10 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in human breast carcinoma (PMID: 29375730). The section was incubated with ab271032 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse cardiac muscle. The section was incubated with ab271032 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in rat kidney. The section was incubated with ab271032 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green) Confocal image showing cytoplasmic and nuclear staining in HeLa cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic and nuclear staining in F9 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MOLT-4 cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing weak staining in MOLT-4 cells.Low expression: MOLT-4 (PMID: 15567145) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Flow Cytometry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast, Left) and HeLa (Human cervix adenocarcinoma epithelial cell, Right) cells labelling HUWE1/Mule with ab271032 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Low expression: MOLT-4 (PMID: 15567145).
Flow Cytometry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (Mouse embryonal carcinoma epithelial cell) cells labelling HUWE1/Mule with ab271032 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

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

Isotype control

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications