Description
Glutamine Assay Kit (Colorimetric) (BA0113) (BA0113)
The Glutamine Assay Kit (Colorimetric) (SKU: BA0113) provides a simple, direct and automation-ready colorimetric method for measuring glutamine. Glutamine is an amino acid that plays major roles in protein synthesis, acid-base balance and anabolic processes and serves as a cellular energy and carbon source. The assay is based on hydrolysis of glutamine to glutamate followed by colorimetric determination of the product. The intensity of the product colour, measured at 565 nm, is proportional to the glutamine concentration in the sample. The room-temperature procedure requires no 37C heater and is readily automated for high-throughput work.
| Product Name: | Glutamine Assay Kit (Colorimetric) (BA0113) |
| SKU: | BA0113 |
| Detection Method: | Colorimetric (565 nm) |
| Detection Range: | 0.023 - 2 mM glutamine |
| Sample Type: | Serum, plasma, urine, tissue extracts and cell culture samples |
| Species Reactivity: | All |
| Assay Time: | 40 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A colorimetric assay for glutamine in biological samples using a single working reagent, a 40-minute incubation and a stop reagent. Samples containing glutamate require a sample blank.
- Sensitive and accurate using a 20 uL sample, with a linear range of 0.023 - 2 mM glutamine
- Convenient single working reagent, 40-minute room-temperature incubation, no 37C heater required
- High-throughput and readily automated
- Direct assay of glutamine in serum, plasma, urine, tissue extracts and cell culture samples
- Studying the effects of drugs on glutamine metabolism
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 | Standard Curve: prepare 2.0 mM glutamine Premix by mixing 5 uL of 100 mM Standard with 245 uL distilled water, then dilute per the table. Transfer 20 uL of each standard into a clear flat-bottom 96-well plate. |
| 2 | Samples: add 20 uL of each sample to separate wells. If a sample is known to contain glutamate, add a second aliquot as a sample blank. |
| 3 | Reaction: spin enzyme and reagent tubes briefly. For each standard and sample well, mix 65 uL Assay Buffer, 1 uL Enzyme A, 1 uL Enzyme B, 2.5 uL NAD and 14 uL MTT. For sample blanks omit Enzyme A. Add 80 uL of the appropriate reagent per well and tap to mix. |
| 4 | Incubate 40 minutes at room temperature, add 100 uL Stop Reagent to each well and read OD at 565 nm (520-600 nm). |
Subtract the water blank OD from the standard values and plot delta-OD against standard concentration. [Glutamine] = (ODSAMPLE - ODBLANK) / Slope x n (mM), where ODBLANK is water (or the sample blank if the sample contains glutamate) and n is the dilution factor. If the calculated concentration exceeds 2 mM, dilute in distilled water and repeat. 1 mM glutamine equals 14.6 mg/dL or 146 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 15 mL | -20C |
| NAD Solution | 1 mL | -20C |
| Enzyme A | 120 uL | -20C |
| MTT Solution | 2 x 1.5 mL | -20C |
| Enzyme B | 220 uL | -20C |
| Stop Reagent | 25 mL | -20C |
| Standard (100 mM Glutamine) | 400 uL | -20C |
