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The assay of Tissue Glycogen

The assay of Tissue Glycogen. Glycogen is a polysaccharide of glucose which functions as the secondary short term energy storage in animal cells. It is made primarily by the liver and the muscles. *Principle. Glycogen is released from the tissue by heating with strong alkali

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The assay of Tissue Glycogen

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  1. The assay of Tissue Glycogen • Glycogen is a polysaccharide of glucose which functions as the secondary short term energy storage in animal cells. • It is made primarily by the liver and the muscles.

  2. *Principle • Glycogen is released from the tissue by heating with strong alkali • precipitated on the addition of ethanol. • Sodium sulphate is added as a co precipitant to give a quantitative yield of glycogen. • The polysaccharide is then hydrolyzed in acid and the glucose released is estimated.

  3. 60 5 55 10 50 45 15 40 20 35 25 30 1. Isolation of Glycogen 20 min. 5 min. Occasional Shaking 5ml Ethanol 95% 1.5 g 0.2ml saturated Na2SO4 Occasional Shaking Stand on ice 5min. 2ml KOH Boiling water bath Centrifuge 10min at 4500rpm

  4. 1. Isolation of Glycogen Warm Mix Keep it for the next week Complet to 10ml with D.W. 5ml D.W. ppt

  5. 60 5 55 10 50 45 15 40 20 35 25 30 2. Hydrolysis & Estimation of Glycogen 1drop phenol red 0.5 mol/L NaOH Dilute to 5ml with D.W. 1 hour 1ml HCL 1ml sample Boiling water bath

  6. 60 5 55 10 50 45 15 40 20 35 25 30 3,5-dinitrosalicylic acid Method COOL 5min. 7ml D.W. 2ml DNS 1ml sample Boiling water bath Read the Optical Density at 540 nm

  7. Results Samar A. Damiati

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