穀胱甘肽熒光檢測試劑盒

穀胱甘肽熒光檢測試劑盒可定量測量各種樣品中存在的還原態穀胱甘肽 (GSH) 和氧化態穀胱甘肽 (GSSG)。驗證的樣本類型包括：人類全血；血清; EDTA和肝素血漿；尿; 和分離的紅細胞。大多數細胞裂解物和組織勻漿也是相容的。

Background Glutathione, L-γ-glutamyl-L-cysteinylglycine or GSH, is the highest concentration non-protein thiol in mammalian cells. GSH plays a key role in many biological processes, including the synthesis of proteins and DNA, the transport of amino acids, and the protec¬tion of cells against oxidation. Due to the genetically conserved molecular processes by which cells die, intracellular levels of GSH can favor a cell death pathway via apoptosis (adequate intracellular GSH stores) or via necrosis or autophagy (depleted intracellular GSH stores). Harmful cellular levels of hydrogen peroxide are minimized by the enzyme glutathione peroxidase (GP) using GSH as a reductant. ICT’s Glutathione Fluorescent Detection Kit is designed to quantitatively measure reduced state glutathione (GSH), and oxidized state glutathione (GSSG) present in a variety of samples.

Target Glutathione and Oxidized Glutathione

Method of Analysis Absorbance Plate Reader

Sample Type Whole blood, serum, plasma, erythrocytes, urine, cell lysates, and tissue samples

Storage 2-8°C

Country of Origin United States

Protocols

1. Use plate layout template to aid in proper sample and standard identification.

2. Pipet 50 µL SSA-treated samples, standards, or control into plate wells.

3. Pipet 50 µL Sample Diluent into zero wells.

4. Add 25 µL Fluorescent GSH Detection Substrate to each well using a repeater pipet.

5. Gently tap plate sides to ensure adequate mixing of the reagents.

6. Incubate 15 minutes at room temperature.

7. Read the fluorescent emission at 510 nm with excitation at 370-410 nm. This data will be used to determine Free GSH concentration.

8. Add 25 µL Reaction Mixture to each well using a repeater pipet.

9. Gently tap plate sides to ensure adequate mixing of the reagents.

10. Incubate 15 minutes at room temperature.

11. Read the fluorescence with excitation at 370-410 nm and emission at 510 nm. This data will be used to determine Total GSH concentration.

Kit Contents

Kit #9133: 1 black half-area polystyrene 96-round well non-binding surface treated microtiter plate, #269 Glutathione Standard, 250 µM, 100 µL, #6610 Fluorescent GSH Detection Substrate, 2 vials, #6612 Dry DMSO, 4 mL, #6614 Assay Buffer, 1X, 60 mL, #6616 NADPH Concentrate, 300 µL, #6618 Glutathione Reductase Concentrate, 300 µL, #6620 Oxidized Glutathione Control, 300 µL, #6623 Kit Manual