Under the presence of concentrated sulfuric acid, glycogen can be dehydrated to furfural derivatives. Furfural derivatives can form blue compound with anthracenone. The concentration of the compound can be measured by colorimetric quantification at 620 nm with glucose standard buffer of same treatment. Glycogen is quite stable in concentrated alkali solution. Heating the tissue sample in concentrated alkali solution before color development will remove other components and keep the glycogen.
Glycogen produces glucose under the action of starch glycosidase, and glucose is catalyzed by glucose oxidase to produce hydrogen peroxide. In the presence of the peroxidase, hydrogen peroxide be oxidized to produce the fluorescence substrate. The fluorescence intensity at the excitation wavelength of 535 nm and emission wavelength of 587 nm is proportional to the glycogen content.
Glycolate oxidase catalyzes sodium glycolate substrate to form glyoxylic acid, which reacts with phenylhydrazine hydrochloride to form phenylhydrazone glyoxalate. The substance has an absorption peak at 324 nm, and its OD value is proportional to the concentration of phenylhydrazone glyoxalate in a certain range, and the amount of phenylhydrazone generated reflects the activity of glycolate oxidase.
Glycolysis is an important ATP generation pathway in eukaryotic cells. Glycolytic stress testing can directly measure the extracellular acidification rate (ECAR) to evaluate the changes of glycolytic process in living cells, and can obtain key parameters for a comprehensive assessment of glycolytic pathway, including: glycolysis, glycolytic capacity, glycolytic reserve and non-glycolytic acidification. This kit uses 2-deoxy-D-glucose (2-DG) and oligomycin to facilitate in-depth analysis of cellular glycolytic flux: 2-DG quantifies nonglycolytic extracellular acidification (ECA) by blocking glycolysis through competitive hexokinase inhibition; Oligomycin inhibits ATP synthetase and therefore prevents aerobic ATP production, forcing the cell to increase glycolytic flux to meet ATP demand. Such compensatory glycolysis levels achievable under the test conditions could reveal glycolytic perturbations that are not apparent under basal conditions and help assess glycolytic capacity with limited ATP demand.
GSP with ketoamine structure is able to react with tetrazole blue under alkaline conditions to form purplish red product, which has a characteristic absorption peak at 530 nm, the concentration of GSP can be calculated by measuring the OD value.
Granule-bound Starch Synthase (GBSS) catalyzes the reaction between ADPG and the primer, transferring glucose molecules to the primer and generating ADP, which in turn catalyzes the reduction of NADP+ to NADPH. The amount of NADPH produced was proportional to the amount of ADP produced in the previous reaction step, and GBSS activity was calculated by measuring the increase in NADPH at 340 nm.
H2S can react with acetate solution to form ZnS which can be dissolved in alkaline solution. In the presence of Fe3+, methylene blue can be formed. Methylene blue has a maximum absorption peak at 665 nm. H2S content can be calculated indirectly by measuring the OD value at 665 nm.
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