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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

Reviews (1)Q&A (1)References (18)
Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Immunoprecipitation - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • OI-RD Scanning - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
  • Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7154(B)] to MGEA5/OGA
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-MGEA5/OGA antibody [EPR7154(B)]
    See all MGEA5/OGA primary antibodies

  • Description

    Rabbit monoclonal [EPR7154(B)] to MGEA5/OGA

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human MGEA5/OGA. The exact sequence is proprietary.
    (Peptide available as ab155521)

  • Positive control

    • WB: HEK-293T, HeLa, BxPC-3, JAR and fetal brain lysates. IHC-P: Human kidney and colon tissue. IP: HeLa 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124807 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)
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).Can be blocked with MGEA5/OGA peptide (ab155521).

For unpurified use at 1/10000 - 1/5000.
IP
1/30.

For unpurified use at 1/100 - 1/500.
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified use at 1/100 - 1/250.

See IHC antigen retrieval protocols.
Notes
WB
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).Can be blocked with MGEA5/OGA peptide (ab155521).

For unpurified use at 1/10000 - 1/5000.
IP
1/30.

For unpurified use at 1/100 - 1/500.
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified use at 1/100 - 1/250.

See IHC antigen retrieval protocols.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • Function

    Isoform 1: Cleaves GlcNAc but not GalNAc from O-glycosylated proteins. Can use p-nitrophenyl-beta-GlcNAc and 4-methylumbelliferone-GlcNAc as substrates but not p-nitrophenyl-beta-GalNAc or p-nitrophenyl-alpha-GlcNAc (in vitro) (PubMed:11148210). Does not bind acetyl-CoA and does not have histone acetyltransferase activity (PubMed:24088714).
    Isoform 3: Cleaves GlcNAc but not GalNAc from O-glycosylated proteins. Can use p-nitrophenyl-beta-GlcNAc as substrate but not p-nitrophenyl-beta-GalNAc or p-nitrophenyl-alpha-GlcNAc (in vitro), but has about six times lower specific activity than isoform 1.

  • Tissue specificity

    Ubiquitous. Shows highest expression in the brain, placenta and pancreas.

  • Sequence similarities

    Belongs to the glycosyl hydrolase 84 family.

  • Post-translational
    modifications

    Proteolytically cleaved by caspase-3 during apoptosis. The fragments interact with each other; cleavage does not decrease enzyme activity.

  • Cellular localization

    Cytoplasm and Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Beta-hexosaminidase antibody
      • Beta-N-acetylglucosaminidase antibody
      • Beta-N-acetylhexosaminidase antibody
      • Bifunctional protein NCOAT antibody
      • HEXC antibody
      • Hyaluronidase in meningioma antibody
      • KIAA0679 antibody
      • MEA5 antibody
      • Meningioma expressed antigen 5 antibody
      • Meningioma-expressed antigen 5 antibody
      • MGEA 5 antibody
      • Mgea5 antibody
      • N-acetyl-beta-D-glucosaminidase antibody
      • N-acetyl-beta-glucosaminidase antibody
      • NCOAT antibody
      • Nuclear cytoplasmic O GlcNAcase and acetyltransferase antibody
      • Nuclear cytoplasmic O-GlcNAcase and acetyltransferase antibody
      • OGA antibody
      • OGA_HUMAN antibody
      • Protein O-GlcNAcase antibody
      see all

    Images

    • Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      All lanes : Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : MGEA5 knockout HeLa cell lysate
      Lane 3 : JAR cell lysate
      Lane 4 : HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 102 kDa
      Observed band size: 130 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab124807 observed at 130 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab124807 Anti-MGEA5/OGA antibody [EPR7154(B)] was shown to specifically react with MGEA5/OGA in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265821 (knockout cell lysate ab257263) was used. Wild-type and MGEA5/OGA knockout samples were subjected to SDS-PAGE. ab124807 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling MGEA5/OGA with purified ab124807 at 1:50 dilution (12.56 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807) at 1/5000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 20ug at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 102 kDa
      Observed band size: 130 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Immunoprecipitation - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

      ab124807 (purified) at 1:30 dilution (2µg) immunoprecipitating MGEA5/OGA in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab124807 & HeLa whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124807 in HeLa whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Western blot - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      All lanes : Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807) at 1/10000 dilution

      Lane 1 : HeLa lysate
      Lane 2 : BxPC-3 lysate
      Lane 3 : JAR lysate
      Lane 4 : Fetal brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 102 kDa
      Observed band size: 130 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

      ab124807, at 1/100 dilution, staining MGEA5/OGA in paraffin-embedded Human kidney tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      OI-RD Scanning - Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)
      Anti-MGEA5/OGA antibody [EPR7154(B)] (ab124807)

    References (18)

    Publishing research using ab124807? Please let us know so that we can cite the reference in this datasheet.

    ab124807 has been referenced in 18 publications.

    • Oliveira-Nunes MC  et al. O-GlcNAcylation protein disruption by Thiamet G promotes changes on the GBM U87-MG cells secretome molecular signature. Clin Proteomics 18:14 (2021). PubMed: 33902430
    • Li C  et al. Hypertonic stress modulates eNOS function through O-GlcNAc modification at Thr-866. Sci Rep 11:11272 (2021). PubMed: 34050207
    • Seo J  et al. O-Linked N-Acetylglucosamine Modification of Mitochondrial Antiviral Signaling Protein Regulates Antiviral Signaling by Modulating Its Activity. Front Immunol 11:589259 (2020). PubMed: 33603735
    • Kim YJ  et al. O-GlcNAc stabilizes SMAD4 by inhibiting GSK-3ß-mediated proteasomal degradation. Sci Rep 10:19908 (2020). PubMed: 33199824
    • Yang Y  et al. O-GlcNAc transferase inhibits visceral fat lipolysis and promotes diet-induced obesity. Nat Commun 11:181 (2020). PubMed: 31924761
    View all Publications for this product

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-MGEA5/OGA antibody [EPR7154(B)]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse embryonic stem cell)
    Gel Running Conditions
    Reduced Denaturing (4-12%, MOPS buffer)
    Loading amount
    20 µg
    Specification
    Mouse embryonic stem cell
    Blocking step
    Milk as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted May 07 2012

    Question

    abcam,

    Hello, abcam.

    I ordered a antibody(ab124807) from your company recently, and I got it several weeks ago.

    I have a question about the product.

    I tested it by western blotting, and it shows several bands.

    So I want to check whether those are specific or not.

    Because it is known that m/w of OGA's splice variant form is almost 70 kDa, and that of caspase-cleaved OGA is around 65 kDa.

    I want to know the exact epitope(peptide region) of this ab.(ab124807).

    See the attached files, please.

    Thank you.

    Read More
    Answer

    As discussed in my previous email, I have now arranged for the immunizing peptide to be available from our catalogue. This can be viewed under the catalogue number ab155521:

    https://www.abcam.com/ab155521

    I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

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