Recombinant
RabMAb

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

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
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: HeLa, BxPC-3, JAR and fetal brain lysates; IHC-P: Human kidney and colon tissue IP: HeLa cells.
  • General notes

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

     

    Previously labelled as MGEA5

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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 1/5000. 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.
  • 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

    • 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 Perform heat mediated antigen retrieval 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.

    • 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?

    • 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.

    • 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?

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

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Huang R  et al. Increased Ratio of Global O-GlcNAcylation to Tau Phosphorylation at Thr212 Site Is Associated With Better Memory Function in Patients With Type 2 Diabetes. Front Physiol 10:110 (2019). Read more (PubMed: 30837891) »
    • Shi H  et al. Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity. Mol Metab N/A:N/A (2018). Read more (PubMed: 29525407) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    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
    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

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    Verified customer

    Submitted May 07 2012

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