Recombinant Anti-MGEA5/OGA antibody [EPR7154(B)] - BSA and Azide free (ab227989)
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 - BSA and Azide free
- Suitable for: IP, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MGEA5/OGA antibody [EPR7154(B)] - BSA and Azide free
See all MGEA5/OGA primary antibodies -
Description
Rabbit monoclonal [EPR7154(B)] to MGEA5/OGA - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab155521) -
Positive control
- IHC-P: Human kidney and colon tissues. IP: HeLa cells. WB: HeLa, JAR, and HEK-293T cell lysates.
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General notes
ab227989 is the carrier-free version of ab124807.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 9.80 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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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 ab227989 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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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa).
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Notes |
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IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 130 kDa (predicted molecular weight: 102 kDa). |
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?This data was developed using the same antibody clone in a different buffer formulation (ab124807).
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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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. -
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124807).
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This IHC data was generated using the same anti-MGEA5/OGA antibody clone, EPR7154(B), in a different buffer formulation (cat# ab124807).
ab124807, at 1/100 dilution, staining MGEA5 in paraffin-embedded Human kidney tissue by Immunohistochemistry.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab227989 has been referenced in 1 publication.
- Park HC et al. Urinary N-acetyl-ß-D glucosaminidase as a surrogate marker for renal function in autosomal dominant polycystic kidney disease: 1 year prospective cohort study. BMC Nephrol 13:93 (2012). IHC-Fr ; Human . PubMed: 22935351