(4 references)-
- 抗原 See all Glutamate products
- Glutamate
- 应用范围
- Biochemical Assay (BCA)
- 样品类型
- Serum, Plasma, Tissue Homogenate, Food
- 特异性
- 50 μM
- 产品特性
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Sensitive and accurate. Detection limit of 50 µM, linearity up to 2.5 mM glutamate in 96-well plate assay.
Convenient. The procedure involves adding a single working reagent, and reading the optical density at time zero and at 30 min at room temperature. No 37°C heater is needed.
High-throughput. Can be readily automated as a high-throughput 96- well plate assay for thousands of samples per day. - 组件
- Assay Buffer: 10 mL. NAD Solution: 1 mL. Enzyme Mix: 120 µL. MTT Solution: 1.5 mL. Standard: 1 mL 100 mM Glutamate.
- 试剂未包括
- Pipeting (multi-channel) devices. Clear-bottom 96-well plates (e.g. Corning Costar) and plate reader.
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- 应用备注
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Direct Assays: glutamate in serum, plasma, tissue extracts and food extract samples.
Drug Discovery/Pharmacology: effects of drugs on glutamate levels. - 说明
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1. This assay is based on an enzyme-catalyzed kinetic reaction. Addition of Working Reagent should be quick and mixing should be brief but thorough. Use of multi-channel pipettor is recommended.
2. The following substances interfere and should be avoided in sample preparation: EDTA (>0.5 mM), ascorbic acid, SDS (>0.2%), sodium azide, NP-40 (>1%) and Tween-20 (>1%). - 实验流程
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1. Calibration Curve. Prepare 600 µL
2.5 mM Glutamate Premix by mixing 15 µL 100 mM Standard and 585 µL distilled water. Transfer 20 µL standards into wells of a clear bottom 96-well plate. Samples: add 20 µL sample per well in separate wells. IMPORTANT: Serum and tissue extract samples require a sample blank.
2. Reaction. Add 80 µL Working Reagent (or Blank Working Reagent where appropriate) per reaction well quickly. Tap plate to mix briefly and thoroughly.
3. Read optical density (OD0) for time zero at 565 nm (520-600nm) and OD30 after a 30-min incubation at room temperature. - 试剂准备
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Spin the Enzyme Mix tube briefly before pipetting. For each well of reaction, prepare Working Reagent by mixing 60 µL Assay Buffer, 1 µL Enzyme Mix, 5 µL NAD and 14 µL MTT. Fresh reconstitution is recommended. Where a sample blank in required, prepare a Blank Working Reagent by mixing 60 µL Assay Buffer, 5 µL NAD and 14 µL MTT (i.e. No Enzyme Mix).
- 限制
- 仅限研究用
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- 储存条件
- -20 °C
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: "Native presynaptic metabotropic glutamate receptor 4 (mGluR4) interacts with exocytosis proteins in rat cerebellum." in: The Journal of biological chemistry, Vol. 287, Issue 24, pp. 20176-86, (2012) (PubMed).
: "Additive effect of tetramethylpyrazine and deferoxamine in the treatment of spinal cord injury caused by aortic cross-clamping in rats." in: Spinal cord, Vol. 49, Issue 2, pp. 302-6, (2011) (PubMed).
: "Manipulating the permeation of charged compounds through the MscL nanovalve." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 25, Issue 1, pp. 428-34, (2011) (PubMed).
: "Oxygen-inducible glutamate oxaloacetate transaminase as protective switch transforming neurotoxic glutamate to metabolic fuel during acute ischemic stroke." in: Antioxidants & redox signaling, Vol. 14, Issue 10, pp. 1777-85, (2011) (PubMed).
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: "Native presynaptic metabotropic glutamate receptor 4 (mGluR4) interacts with exocytosis proteins in rat cerebellum." in: The Journal of biological chemistry, Vol. 287, Issue 24, pp. 20176-86, (2012) (PubMed).
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- 抗原
- Glutamate
- Abstract
- Glutamate 产品
- 物质类
- Amino Acid
- 背景
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Quantitative determination of glutamate by colorimetric (565nm) method.
Procedure: 30 min.
Glutamate is an important chemical in general metabolism. It is also a crucial mammalian neurotransmitter that is believed to be involved in a number of neurological and psychiatric disorders such as lateral sclerosis, lathyrism, autism and Alzheimer's disease. Glutamate is also widely used as a flavor enhancer in the food industry. Simple, direct and automation-ready procedures for measuring glutamate concentration are very desirable. This glutamate assay kit is based on glutamate dehydrogenase catalyzed oxidation of glutamate, in which the formed NADH reduces a formazan (MTT) Reagent. The intensity of the product color, measured at 565 nm, is proportionate to the glutamate concentration in the sample.
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