Description
O-GlcNAcase (OGA/MGEA5) Activity Assay Kit
The Assay Genie O-GlcNAcase (OGA) Activity Assay Kit measures O-GlcNAcase (OGA, also known as MGEA5 or NCOAT, EC 3.2.1.169) activity in cell and tissue lysates, serum and plasma. OGA removes O-linked β-N-acetylglucosamine (O-GlcNAc) from serine and threonine residues of nuclear and cytoplasmic proteins and, with O-GlcNAc transferase (OGT), controls dynamic O-GlcNAc cycling linked to diabetes, cardiovascular disease, cancer and neurodegeneration.
OGA cleaves the chromogenic substrate p-nitrophenyl-N-acetyl-β-D-glucosaminide; adding NaOH stops the reaction and develops yellow p-nitrophenol, read at 405 nm. A dedicated Control Solution corrects for background in coloured samples such as serum and plasma, and activity is calculated directly in IU/L.
| Product Name: | O-GlcNAcase Assay Kit |
| SKU: | BR00012 |
| Kit Size: | 100 wells |
| Detection Method: | Colorimetric (p-nitrophenol release, OD 405 nm) |
| Sample Types: | Cell and tissue lysates; serum and plasma (colour-control format) |
| Recommended Protein: | 0.5–4 mg/ml |
| Incubation Time: | 30 or 60 min at 37 °C |
| Units: | IU/L (µmol/L/min) |
| Equipment Required: | Microplate reader (405 nm) |
| Storage: | -20 °C (see component storage) |
| Intended Use: | For research use only (RUO). Not for diagnostic procedures. |
- Compatible with serum and plasma via a colour-control format
- Chromogenic pNP-GlcNAc substrate with simple NaOH stop
- Direct activity calculation in IU/L, no standard curve required
- Results in 30–60 min
- Includes 10x Cell Lysis Solution
- OGA hydrolyses p-nitrophenyl-N-acetyl-β-D-glucosaminide, releasing p-nitrophenol.
- Addition of 1 N NaOH stops the reaction and develops the yellow p-nitrophenolate, read at 405 nm.
- Clear samples: IU/L = ΔOD × 21.6 (30 min) or × 10.8 (60 min). Coloured samples: ΔOD × 40.12 (30 min) or × 20.06 (60 min).
- Cells: ~105 washed cells in 50–100 µl ice-cold 1x Cell Lysis Solution, 5 min on ice.
- Tissue: ~5 mg tissue homogenised in 0.5 ml ice-cold 1x Cell Lysis Solution.
- Centrifuge at ~14,000 rpm for 5 min at 4 °C; protein 0.5–4 mg/ml.
- Serum/plasma: use directly with the coloured-sample procedure.
| Component | Size | Storage |
| OGA Assay Solution | 10 ml | -20 °C |
| OGA Control Solution | 10 ml | 4 °C |
| 10x Cell Lysis Solution | 25 ml | Room temperature |
- O-GlcNAc cycling and nutrient-sensing signalling
- Diabetes and insulin resistance research
- Neurodegeneration and tau O-GlcNAcylation studies
- Cancer biology and screening of OGA inhibitors
Can I use serum or plasma with the OGA kit?
Yes. Coloured samples such as serum and plasma are run with the supplied OGA Control Solution to correct for background absorbance.
How is OGA activity calculated?
For clear samples, ΔOD at 405 nm × 21.6 (30 min) or × 10.8 (60 min) gives IU/L; coloured samples use the factors 40.12 and 20.06.
