产品说明
DescriptionQ & AManualS.D.SProduct DescriptionGlutamine, a source of α-ketoglutarate (one of the tricarboxylic acid (TCA) cycle intermediates), is an important substance used for energy production and the syntheses of nucleic acids and other amino acids. Glutaminolysis, the process by which glutamine as a substrate is converted to α-ketoglutarate, is upregulated in cancer cells. A study reports that glutaminolysis has largely contributed to scavenging of reactive oxygen species and reduction of oxidized glutathione.The Glutamine Assay Kit-WST can quantify glutamine, a substrate of energy metabolism. Our Glutamine Assay Kit-WST is designed to quantify glutamine as a metabolite. This kit allows you to quantify glutamine present in culture medium or intracellular glutamine by WST reduction reaction. The lowest concentration of glutamine which can be quantified is 5 μmol/l. This kit can be used with 96-well microplates, which makes it possible to analyze multiple specimens.PrincipleThe kit is used to detect Glutamine in cell culture medium by measuring the absorbance of WST formazan produced according to quantities of glutamine. The kit includes a “Glutamine Standard” which can be used to quantify the concentration of lactate found within samples by creating standard curve.Simple Assay Procedure*The glutamine concentration can be calculated by using the following equation:glutamine (mmol/l) = (Glutaminase solution (+))－(Glutaminase solution (-))Standard CurveStandard curve can be prepared by using Glutamine Standard included in the kit. The concentration of Glutamine can be measured. The concentration of glutamine can be evaluated by diluting the samples if the concentration is over 0.5 mmol/l.Example of Measurements of Glutamine/Glutamate levelA549 cells were seeded on 6-well plates. Glutamine concentrations in the culture supernatant over incubation time were measured using the Glutamine Assay Kit-WST, and changes in glutamate concentration were measured using the Glutamate Assay Kit-WST. As a result, upregulated glutaminolysis led to a time-dependent decrease in glutamine concentrations and a time-dependent increase in glutamate concentrations in culture medium.How many samples can I measure?The number of samples that can be recorded when the standard curve and sample is measured in triplicates.100 testsSample 12 samples*(measured in triplicate – See picture below)A plate layout for Glutamate standard solution and samplePrepare a test sample to be assayed. Please note that a minimum of 240 μl of sample (e.g., supernatant, lysate) is needed for each test sampleThe glutamine concentration can be calculated by using the following equation:glutamine (mmol/l) = (Glutaminase solution (+))－(Glutaminase solution (-))How stable is Working solution?You can’t store the Working solution. Please prepare the Working solution prior to use. Please protect from light because the Working solution is light-sensitive.Can I quantify D-Glutamine levels?No, the kit is used for the quantification of L-Glutamine levels.Can I measure samples that contain a reducing agent?You can’t accurately measure Glutamine levels if your samples contain a reducing agent. WST dye interferes with the reducing agent and can cause errors in observations. If you add a testing material that has reducing properties, please prepare a few wells that only contains the medium (no cells) and testing material for background control.Our samples did not change in color, are there any reasons for this?Glutamine level found within the sample may be lower than the detection limit (5 µmol/l) that can be measured using this kit.If you dilute the sample, lactate level present in the diluted sample may be lower than 5 µmol/l.Please lower the dilution ratio and adjust the concentration of the sample over the detection limit that was measured using this kit.Related Categories Cell Proliferation / Cytotoxicity Intracellular Fluorescent Probes Cellular Metabolism
Dojindo细胞分析
细胞活力和细胞毒性测定用于药物筛选和化学物质的细胞毒性测试。Dojindo开发了高度水溶性的四唑盐，称为WST。WST-8是高度稳定的WST，用于Cell Counting Kit-8（CCK-8）。由于WST-8甲maz是水溶性的，因此不会形成晶体。因此，不需要诸如MTT测定的增溶过程。此外，CCK-8的检测灵敏度高于其他四唑盐，例如MTT，XTT，MTS或WST-1。
细胞增殖/细胞毒性转染细胞染色细胞内荧光探针细菌染色微生物活力测定干细胞分化SPiDER-ßGal线粒体检测细胞代谢
| 应用 | 产品展示 |
| 细胞生长检测，药物筛选，比色/荧光检测 | 细胞计数试剂盒-8 细胞计数试剂盒8 + 96孔有机硅定向剂 细胞计数试剂盒-F 细胞毒性LDH检测试剂盒 -WST 96孔有机硅定向剂 MTT |
| 了解检测机制的差异： | 点击这里 |
| 细胞周期分析 | 细胞周期测定溶液深红色 细胞周期测定溶液蓝色 |
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