{"product_id":"ua-glo-glycogen-luminescence-detection-kit-ua079050","title":"UA-Glo® Glycogen Luminescence Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSynonyms\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eUA-Glo® 糖原发光法检测试剂盒\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eDry ice transportation. Store protected from light at -80°C, validity period as indicated on the reagent bottle label. After initial use, it is recommended to aliquot the reagent as needed and store protected from light at -80°C, preferably in single-use aliquots. The reductase substrate and luciferin detection solution should not undergo more than 3 freeze-thaw cycles. 0.3N HCl and Tris buffer can be stored at 4°C.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e\u003cspan\u003eThe UA-Glo® Glycogen Luminescence Assay Kit detects glycogen content in samples through a two-step reaction. The first step involves the digestion of glycogen into glucose by glucoamylase (1,4-α-D-Glucan glucohydrolase). In the second step, the glucose content is measured via a coupled reaction involving glucose dehydrogenase, reductase, and luciferase. Glucose dehydrogenase acts on glucose to reduce NADP to NADPH, while reductase utilizes NADPH to reduce the reductase substrate (also a luciferin precursor) into luciferin. Finally, luciferin is catalyzed by luciferase to produce luminescence, the intensity of which is directly proportional to the glucose concentration.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eThe UA-Glo® Glycogen Luminescence Assay Kit is a homogeneous detection reagent. Its simple two-step procedure minimizes experimental errors caused by multiple pipetting steps. This kit offers high signal-to-noise ratio, excellent reproducibility, and stability, making it suitable for batch processing of various sample types.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eComponents\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003eThe components and specifications of the UA-Glo® Glycogen Luminescence Assay Kit are as follows. The detectable 96-well plate reactions are calculated based on adding 25 μL of sample, 25 μL of glucoamylase digestion solution, and 50 μL of glucose detection reagent per well.\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSpecification\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e96-Well Plate Reactions\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e5ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucoamylase: 15 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucoamylase Buffer: 3 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLuciferin Detection Reagent: 5 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eReductase Substrate: 25 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlycogen (2mg\/mL): 50 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucose (10mM): 50 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0.3N HCl: 15 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eTris Buffer: 15 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e100\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003e50ml\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucoamylase: 150 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucoamylase Buffer: 30 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eLuciferin Detection Reagent: 50 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eReductase Substrate: 250 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlycogen (2mg\/mL): 50 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eGlucose (10mM): 50 μL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e0.3N HCl: 15 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eTris Buffer: 15 mL\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003e1,000\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp\u003e# Translation Result:\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eThe UA-Glo® Glycogen Assay Kit detects glycogen content through glucose amylase digestion of glycogen into glucose, followed by coupled reactions involving glucose dehydrogenase, reductase, and luciferase to measure glucose levels. The luciferase reaction offers extremely high sensitivity, but parameters such as pH, temperature, and experimental system setup significantly influence the results. Under most experimental conditions, direct comparison of absolute readings between samples is not recommended. It is advised to include the same reference compounds and solvent vehicle controls on each assay plate, with sample readings normalized to these controls before data processing and comparison. Alternatively, glycogen standards processed in the same manner as samples may be included on each plate to quantify glycogen content.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.1\u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e Sample Processing\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan\u003eThe UA-Glo® Glycogen Assay Kit can be used to measure glycogen concentration in cell lysates and tissue homogenates. The linear detection range for glycogen extends up to \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e20 μg\/mL\u003c\/span\u003e\u003cspan\u003e. If sample glycogen concentration exceeds \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e20 μg\/mL\u003c\/span\u003e\u003cspan\u003e, refer to the table below for appropriate sample processing and dilution, or optimize dilution based on experimental requirements.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eIt is recommended to use the kit-supplied \u003c\/span\u003e\u003cspan\u003e0.3N HCl\u003c\/span\u003e\u003cspan\u003e and \u003c\/span\u003e\u003cspan\u003eTris buffer (450mM Tris, pH 8.0)\u003c\/span\u003e\u003cspan\u003e for sample processing. Acid treatment with \u003c\/span\u003e\u003cspan\u003e0.3N HCl\u003c\/span\u003e\u003cspan\u003e rapidly lyses cells, halts metabolism, inactivates cellular enzymes, and degrades reduced NAD(P)H, preventing interference in subsequent glycogen detection. After acid treatment, adding \u003c\/span\u003e\u003cspan\u003eTris buffer\u003c\/span\u003e\u003cspan\u003e slightly increases pH without negating the acid treatment effect. The combination of \u003c\/span\u003e\u003cspan\u003e0.3N HCl\u003c\/span\u003e\u003cspan\u003e and \u003c\/span\u003e\u003cspan\u003eTris buffer (450mM Tris, pH 8.0)\u003c\/span\u003e\u003cspan\u003e ensures the pH reaches the optimal range (~pH 5) for glucose amylase reaction after adding the digestion buffer. Samples treated with \u003c\/span\u003e\u003cspan\u003e0.3N HCl\u003c\/span\u003e\u003cspan\u003e and \u003c\/span\u003e\u003cspan\u003eTris buffer\u003c\/span\u003e\u003cspan\u003e can be immediately assayed or stored at \u003c\/span\u003e\u003cspan\u003e-20°C\u003c\/span\u003e\u003cspan\u003e or below for future use. For difficult-to-lyse samples (e.g., 3D cultures), \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e0.2% Triton X-100\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e may be added to \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e0.3N HCl\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003e to enhance metabolite release.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003eFor tissue homogenization, use \u003c\/span\u003e\u003cspan\u003e1 volume of PBS\u003c\/span\u003e\u003cspan\u003e mixed with \u003c\/span\u003e\u003cspan\u003e1\/2 volume of 0.3N HCl\u003c\/span\u003e\u003cspan\u003e, followed by addition of \u003c\/span\u003e\u003cspan\u003e1\/2 volume of Tris buffer\u003c\/span\u003e\u003cspan\u003e. Alternatively, homogenize tissue first, then sequentially add \u003c\/span\u003e\u003cspan\u003e0.3N HCl\u003c\/span\u003e\u003cspan\u003e and \u003c\/span\u003e\u003cspan\u003eTris buffer\u003c\/span\u003e\u003cspan\u003e. Tissues with high glycogen content (e.g., liver) require dilution to appropriate concentrations before assay, using a diluent of \u003c\/span\u003e\u003cspan\u003ePBS + 0.3N HCl + Tris buffer\u003c\/span\u003e\u003cspan\u003e in a \u003c\/span\u003e\u003cspan\u003e2:1:1\u003c\/span\u003e\u003cspan\u003e volume ratio.\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eSample\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eGlycogen Concentration\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold\"\u003eProcessing Method\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eCell lysates (intracellular)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"color:#000000\"\u003e1-10 μg\/mL: 60 μL processed sample (20,000 cells)\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eResuspend cells in 1 volume PBS\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eAdd 1\/2 volume 0.3N HCl\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eAdd 1\/2 volume Tris buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan\u003eTissue\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\u003cp style=\"text-align:justify\"\u003e\u003cspan\u003e10 μg\/mL: e.g., 5-15 mg liver tissue homogenized in 1 mL buffer, then diluted 50-100×\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle;border:1px solid #000000\"\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eHomogenize tissue in 1 volume PBS\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eAdd 1\/2 volume 0.3N HCl\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e•\u003c\/span\u003e\u003cspan\u003eAdd 1\/2 volume Tris buffer\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.2 Glycogen Assay Protocol\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.2.1 Sample Glycogen Detection\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eAdd 25 μL of processed sample (from 3.1) per well in a 96-well white opaque-bottom plate. Negative controls should use glycogen\/glucose-free medium or buffer processed identically to samples; positive controls may use glycogen standards similarly processed. Luciferase reactions are temperature-sensitive. Equilibrate reagents and samples to room temperature (22-25°C) and maintain stable temperature (±1°C) during assay.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003eThaw glucose amylase buffer completely at room temperature, mix by inversion, and aliquot as needed. Remaining buffer may be stored at -20°C.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eThaw glucose amylase at room temperature and keep on ice. Prepare 1× digestion solution by diluting 200× stock with glucose amylase buffer, mixing thoroughly. Prepare fresh for each use; do not store. Aliquot remaining enzyme and store at -80°C.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eAdd 25 μL digestion solution per well, mix by plate shaking for 1 min.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eIncubate at room temperature for 60 min.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003eThaw luciferase detection reagent completely, mix by inversion, and equilibrate to room temperature. \u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eAliquot remaining reagent and store at -80°C. Avoid \u0026gt;3 freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7)\u003c\/span\u003e\u003cspan\u003ePrepare glucose detection reagent: Thaw reductase substrate at room temperature, keep on ice. Add to luciferase detection reagent at 200× dilution, mix thoroughly. \u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003eUse immediately to avoid increased background. Prepare fresh; do not store. Aliquot remaining substrate and protect from light at -80°C (≤3 freeze-thaw cycles).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8)\u003c\/span\u003e\u003cspan\u003eAdd 50 μL detection reagent per well. Maintain a 1:1:2 ratio of sample:digestion reagent:detection reagent. For 384-well low-volume plates, use 5 μL sample + 5 μL digestion reagent + 10 μL detection reagent.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9)\u003c\/span\u003e\u003cspan\u003eMix by shaking for 1 min, then incubate in darkness for 60-90 min. \u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eSignal increases linearly until glucose is depleted.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e10)\u003c\/span\u003e\u003cspan\u003eMeasure luminescence using a plate reader.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold\"\u003e3.2.2 Glycogen Detection in Cultured Cell Lysates\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan\u003eSeed cells at appropriate density in 96-well plates (e.g., 5,000-50,000 cells\/well).\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan\u003ePerform experimental treatments as required.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan\u003eAspirate medium and wash cells five times with 200 μL cold PBS to remove residual glucose. Perform washes rapidly to minimize glycogen metabolism changes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan\u003eAdd 30 μL PBS per well.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan\u003eAdd 15 μL 0.3N HCl, mix by shaking for 5 min. Alternatively, premix PBS and 0.3N HCl (2:1) and add 45 μL\/well.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e6)\u003c\/span\u003e\u003cspan\u003eAdd 15 μL Tris buffer (450mM, pH 8.0), mix by shaking for 1 min.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e7)\u003c\/span\u003e\u003cspan\u003eSamples may be stored at ≤-20°C for future analysis.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:justify\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e8)\u003c\/span\u003e\u003cspan\u003eTransfer 25 μL processed samples, controls to white opaque-bottom plates. For samples with high glucose (e.g., tissues), run duplicates: one well with digestion reagent, one with buffer alone. The signal difference represents glycogen content.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e9)\u003c\/span\u003e\u003cspan\u003eProceed with glycogen detection as in \u003c\/span\u003e\u003cspan style=\"font-weight:bold\"\u003e3.2.1\u003c\/span\u003e\u003cspan\u003e.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\u003ch4\u003eGuidelines\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e1) Different batches are not recommended for mixed use\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e2) It is not recommended to change the amount of detection reagents without strict validation\u003c\/p\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e3) For research use only\u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"5ml","offer_id":43096035459147,"sku":"UA079050-5ml","price":620.0,"currency_code":"USD","in_stock":true},{"title":"50ml","offer_id":43096035491915,"sku":"UA079050-50ml","price":3560.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_6878bc91-4224-49ab-8fdd-e798021442db.png?v=1788228139","url":"https:\/\/www.antbioinc.com\/products\/ua-glo-glycogen-luminescence-detection-kit-ua079050","provider":"AntBio","version":"1.0","type":"link"}