Overview

  • Product name

  • Description

    Rabbit polyclonal to MGEA5/OGA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rabbit, Horse, Cow, Dog, Pig, Macaque monkey, Gorilla
  • Immunogen

    Synthetic peptide within Human MGEA5/OGA aa 500-600 conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab123058)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: U87mg; JEG3; PDX1.
  • General notes

    Previously labelled as MGEA5

Properties

Applications

Our Abpromise guarantee covers the use of ab105217 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 Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 103 kDa).
ICC/IF Use a concentration of 1 µg/ml.

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

  • All lanes : Anti-MGEA5/OGA antibody (ab105217) at 1 µg

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : MGEA5 knockout HAP1 whole cell lysate
    Lane 3 : U20S whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 103 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab105217 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab105217 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in MGEA5 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MGEA5 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab105217 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • ab105217 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab105217 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-MGEA5/OGA antibody (ab105217) at 1 µg/ml

    Lane 1 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
    Lane 2 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate
    Lane 3 : PDX1(Pancreatic and duodenal homeobox 1) Whole Cell Lysate
    Lane 4 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 5 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 6 : PDX1(Pancreatic and duodenal homeobox 1) Whole Cell Lysate with Immunising peptide at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 103 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 58 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes

References

This product has been referenced in:

  • Huang L  et al. O-GlcNAc-modified SNAP29 inhibits autophagy-mediated degradation via the disturbed SNAP29-STX17-VAMP8 complex and exacerbates myocardial injury in type I diabetic rats. Int J Mol Med 42:3278-3290 (2018). Read more (PubMed: 30221662) »
See 1 Publication for this product

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