RecombinantRabMAb

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

Submit a review Submit a question
Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Immunocytochemistry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Flow Cytometry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Flow Cytometry - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)
  • Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24361-13] to HUWE1/Mule
  • Suitable for: IHC-P, ICC, Flow Cyt, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • 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.

Properties

  • 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

Applications

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.

Target

  • 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.
  • Information by UniProt

  • Database links

    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

Images

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

Protocols

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

References (0)

Publishing research using ab271032? Please let us know so that we can cite the reference in this datasheet.

ab271032 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab271032.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com