Recombinant 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
Related conjugates and formulations
Overview
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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 asab155521) -
Positive control
- WB: HEK-293T, HeLa, BxPC-3, JAR and fetal brain lysates. IHC-P: Human kidney and colon tissue. IP: HeLa cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 9.80 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7154(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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 |
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WB | (1) |
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).
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. |
Notes |
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WB
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa). 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. |
Target
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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
modificationsProteolytically 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
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Database links
- Entrez Gene: 10724 Human
- Omim: 604039 Human
- SwissProt: O60502 Human
- Unigene: 500842 Human
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Alternative names
- Beta-hexosaminidase antibody
- Beta-N-acetylglucosaminidase antibody
- Beta-N-acetylhexosaminidase antibody
see all
Images
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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.
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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.
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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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab124807 has been referenced in 22 publications.
- Sombutthaweesri T et al. Relationship Between O-GlcNAcase Expression and Prognosis of Patients With Osteosarcoma. Appl Immunohistochem Mol Morphol 30:e1-e10 (2022). PubMed: 34469899
- Wang Y et al. EGF promotes PKM2 O-GlcNAcylation by stimulating O-GlcNAc transferase phosphorylation at Y976 and their subsequent association. J Biol Chem 298:102340 (2022). PubMed: 35931120
- Cao Y et al. O-GlcNAcylation of SIRT1 Protects against Cold Stress-Induced Skeletal Muscle Damage via Amelioration of Mitochondrial Homeostasis. Int J Mol Sci 23:N/A (2022). PubMed: 36498847
- Akinbiyi EO et al. Blocked O-GlcNAc cycling alters mitochondrial morphology, function, and mass. Sci Rep 11:22106 (2021). PubMed: 34764359
- Li C et al. Hypertonic stress modulates eNOS function through O-GlcNAc modification at Thr-866. Sci Rep 11:11272 (2021). PubMed: 34050207