Glutathione Fluorescent Detection Kit

1. Prepare samples; treat with SSA to remove thiol-bearing protein., 2. Prepare standard curve and blanks., 3. If measuring oxidized GSH (GSSG), treat blanks, samples, and standards with 2VP., 4. Add 50 µL blanks, standards, and samples to plate., 5. Add 25 µL Colorimetric GSH Detection Substrate., 6. Add 25 µL Reaction Mixture., 7. Mix., 8. Incubate 20 minutes at room temperature., 9. Read at 405 nm.

Kit 9134:, 5 black half-area polystyrene 96-round Well non-binding surface treated microtiter plate, #269, Glutathione Standard, 250 µM, 300 µL, #6611, Fluorescent GSH Detection Substrate, 4 vials, #6613, Dry DMSO, 20 mL, #6615, Assay Buffer, 1X, 200 mL, #6617, NADPH Concentrate, 1.4 mL, #6619, Glutathione Reductase Concentrate, 1.4 mL, #6621, Oxidized Glutathione Control, 300 µL, #6623, Kit Manual

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.