Human OGA ELISA Kit (ab269546)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.4 ng/ml
- Range: 1.2 ng/ml - 75 ng/ml
- Sample type: Cell culture extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human OGA ELISA Kit
See all MGEA5/OGA kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Extract 8 2.6% Inter-assay Sample n Mean SD CV% Extract 4 2.8% -
Sample type
Cell culture extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
0.4 ng/ml -
Range
1.2 ng/ml - 75 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture extracts 84 81% - 90% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human OGA ELISA Kit (ab269546) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of OGA protein in cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human OGA with 0.4 ng/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Protein O-GlcNAcase (OGA, NAG), also referred to as Beta-N-Acetylhexosaminidase and Meningioma-expressed antigen 5 (MGEA5) is a 916 amino acid enzyme encoded by the gene OGA. OGA catalyzes the hydrolysis of GlcNac (N-acetyl-Beta-D-glucosamine) from O-glycosylated serine or threonine residues to form free N-acetyl-D-glucosamine generally in the cytosol. Thiazoline derivatives of GlcNAc have been found to act as OGA enzyme inhibitors by mimicking the reaction intermediates. OGA has a broad range of proteins that act as substrates, which may modulate downstream effects such as transcription and proteomic activities. OGA plays a role in embryonic development, brain activity, and hormone production.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human OGA Capture Antibody 1 x 600µl 10X Human OGA Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BR 1 x 6ml Human OGA Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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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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Alternative names
- Beta-hexosaminidase
- Beta-N-acetylglucosaminidase
- Beta-N-acetylhexosaminidase
see all -
Database links
- Entrez Gene: 10724 Human
- Omim: 604039 Human
- SwissProt: O60502 Human
- Unigene: 500842 Human
Images
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human OGA standard curve in Sample Diluent 1X Cell Extraction Buffer PTR.The OGA standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native OGA in human SH-SY5Y cell extract based on a 200 µg/mL extract load.The concentrations of OGA were measured in duplicate and interpolated from the OGA standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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Sandwich ELISA - Human OGA ELISA Kit (ab269546)To learn more about the advantages of recombinant antibodies see here.
