Presentation on theme: "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."— Presentation transcript:
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 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.
1.5 g 2ml KOH Boiling water bath min. Occasional Shaking 0.2ml saturated Na 2 SO 4 Occasional Shaking 5ml Ethanol 95% Stand on ice 5min. Centrifuge 10min at 4500rpm 1. Isolation of Glycogen 5 min.
ppt 5ml D.W. Warm Complet to 10ml with D.W. Mix 1. Isolation of Glycogen Keep it for the next week
1ml HCL 2. Hydrolysis & Estimation of Glycogen Boiling water bath hour 1drop phenol red 0.5 mol/L NaOH 1ml sample Dilute to 5ml with D.W.
3,5-dinitrosalicylic acid Method 1ml sample 2ml DNS Boiling water bath min. COOL 7ml D.W. Read the Optical Density at 540 nm