Metabolism Assays
Formate Assay Kit (BA0108)
- SKU:
- BA0108
- Product Type:
- Assay
- Instrument:
- Microplate Reader
- Sample Type:
- Serum, Plasma, Urine
- Research Area:
- TCA Cycle
- Research Area:
- Oxidative Stress
- Research Area:
- Antioxidant
- Research Area:
- Cation and Inorganic Ion
Description
Formate Assay Kit (BA0108) (BA0108)
The Formate Assay Kit (SKU: BA0108) provides a fast, sensitive colorimetric method for determining formate. Formate is the anion of formic acid and a metabolic by-product of formaldehyde and methanol; elevated levels can indicate formaldehyde exposure or methanol poisoning. The assay is based on formate dehydrogenase-catalysed oxidation of formate, which generates NADH that reduces an MTT dye. The intensity of the reduced MTT, measured at 565 nm, is directly proportional to the formate concentration. The room-temperature add-mix-read procedure requires no 37C heater and is readily automated.
| Product Name: | Formate Assay Kit (BA0108) |
| SKU: | BA0108 |
| Detection Method: | Colorimetric (565 nm) |
| Detection Range: | 0.050 - 5 mM formate |
| Sample Type: | Urine, serum and other biological samples |
| Species Reactivity: | All |
| Assay Time: | 60 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A convenient colorimetric assay for formate in biological fluids using only 10 uL of sample and a single working reagent.
- Fast and sensitive using only 10 uL of sample, with a linear range of 0.050 - 5 mM formate
- Convenient single working reagent read after 60 minutes at room temperature, no 37C heater required
- High-throughput add-mix-read format that is readily automated
- Direct assay of formate in biological samples such as urine and serum
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Sample preparation: clear and slightly coloured samples can be assayed directly. Biological fluids such as urine and serum can be assayed after centrifuging to remove particulates; dilute in distilled water as required. |
| 2 | Standards: prepare 200 uL of 1 mM Premix by mixing 10 uL of the 20 mM Standard with 190 uL distilled water, then dilute per the table. |
| 3 | Transfer 10 uL of each standard and 10 uL of each sample into separate wells of a clear flat-bottom 96-well plate. |
| 4 | Prepare Working Reagent per well from 85 uL Assay Buffer, 8 uL MTT/NAD, 1 uL Enzyme A and 1 uL Enzyme B. Add 90 uL to each standard and sample well, tap to mix and incubate 60 minutes at room temperature. |
| 5 | Read optical density at 565 nm (520-600 nm). |
Subtract the blank value from the standard values and plot delta-OD against standard concentration. [Formate] = (ODSAMPLE - ODBLANK) / Slope x n (mM), where n is the sample dilution factor. If a reading exceeds that of the 1 mM standard, dilute in water, repeat and multiply by the dilution factor. 1 mM formate equals 4.5 mg/dL or 45 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20C |
| NAD/MTT | 1 mL | -20C |
| Enzyme A | 120 uL | -20C |
| Enzyme B | 120 uL | -20C |
| Standard (20 mM Formate) | 0.5 mL | -20C |
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