Description
Glucose-6-Phosphatase (G6Pase) Activity Assay Kit
The Assay Genie Glucose-6-Phosphatase (G6Pase) Activity Assay Kit measures G6Pase (EC 3.1.3.9) activity in the endoplasmic reticulum (ER) membrane fraction of cells and tissues, including liver homogenates and microsomes. G6Pase hydrolyses glucose-6-phosphate to free glucose and inorganic phosphate, the terminal step of both gluconeogenesis and glycogenolysis, and is essential for maintaining blood glucose.
Phosphate released by G6Pase is detected as a phosphomolybdate–malachite green complex, read at 630 nm. The kit includes Lysis and Extraction Buffers to isolate the ER membrane fraction, plus TCA to stop the reaction, with a 10-minute enzymatic step and a 5-minute colour development.
| Product Name: | Glucose-6-Phosphatase Assay Kit |
| SKU: | BR00008 |
| Kit Size: | 150 wells |
| Detection Method: | Colorimetric (malachite green phosphate detection, OD 630 nm) |
| Sample Types: | Membrane (ER) fraction of cells and tissues; liver homogenates and microsomes |
| Recommended Protein: | 0.5–2 mg/ml |
| Assay Time: | 10 min at 37 °C + 5 min colour development |
| Readout: | Background-corrected ΔOD; normalise to protein or GAPDH |
| Equipment Required: | Microplate reader (630 nm) |
| Storage: | 4 °C (G6Pase Reaction at -20 °C after reconstitution) |
| Intended Use: | For research use only (RUO). Not for diagnostic procedures. |
- Short 10-minute enzymatic reaction
- Sensitive malachite green phosphate detection
- Two-buffer workflow isolates the ER membrane fraction
- Includes G6Pase Control and TCA Solution for clean background correction
- Lysate supernatant can be kept for hexokinase, GAPDH or LDH assays
- G6Pase hydrolyses glucose-6-phosphate to glucose and inorganic phosphate.
- The reaction is stopped with TCA; released phosphate forms a coloured phosphomolybdate–malachite green complex measured at 630 nm.
- Sample activity = ΔOD (sample) − ΔOD (background), where each ΔOD is reaction minus control.
- Cells: ≥106 cells in ~0.1 ml 1x Lysis Buffer; centrifuge and resuspend the pellet in ~50 µl 1x Extraction Buffer.
- Tissue: ~5 mg tissue homogenised in 0.5 ml 1x Lysis Buffer; resuspend the pellet in ~0.3 ml 1x Extraction Buffer.
- Centrifuge at ~5,000 rpm for 1 min and use the supernatant (0.5–2 mg/ml protein).
| Component | Size | Storage |
| G6Pase Assay Solution | 30 ml | 4 °C |
| 10x Lysis Buffer | 25 ml | 4 °C |
| 10x Extraction Buffer | 20 ml | 4 °C |
| G6Pase Control | 1 ml | 4 °C |
| G6Pase Reaction (lyophilised) | 0.4 ml | -20 °C after reconstitution |
| TCA Solution (10%) | 4 ml | 4 °C |
- Hepatic glucose production, gluconeogenesis and glycogenolysis
- Type 2 diabetes and insulin resistance research
- Glycogen storage disease type I (G6PC/SLC37A4) models
- Liver and kidney ER and microsomal enzyme studies
Why does the G6Pase kit measure the membrane fraction?
G6Pase is an integral ER membrane enzyme. The Extraction Buffer solubilises the membrane pellet left after cytosolic lysis to enrich G6Pase activity.
Can I use liver microsomes?
Yes. The kit is suitable for liver homogenates and microsomes as well as the ER fraction of cells and other tissues.
How many samples can I run?
The kit provides enough assay solution for 150 wells.
