Biochemical Techniques and Instrumentation

Materials

1. Reagent A : Dissolve 2% sodium carbonate (Na₂CO₃) in 0.1N NaOH
2. Reagent B : Dissolve 0.5% copper sulphate (CuSO₄) in 1% sodium potassium tartarate solution
3. Reagent C (Alkaline copper compound): Mix 50 ml of A and 1 ml of B, prior to use
4. Folin–Ciocalteau reagent : Reflux gently a mixture consisting of 100 g sodium tungstate, 25 g sodium molybdate, 100 ml of water, 50 ml of 85% phosphoric acid and 100 ml of conc. HCl in a 1.5 L flask for 10 h. Add 150 g lithium sulphate, 50ml of water and a few drops of bromine water. Boil the mixture for 15 min without condenser to remove excess bromine. Cool, dilute to 1 L and filter. The reagents should have no greenish tinch. (Ready made solution is available)
5. Protein standard solution
   Stock (1mg/ml) : Weigh accurately 50 mg of bovine serum albumin and dissolve in 1N NaOH and make up to 50ml in volumetric flask
   Working (200 m g/ml) : Dilute 10 ml of the stock solution to 50 ml with distilled water in a volumetric flask
6. Phosphate buffer (pH-7.0) : Dissolve 3.403 g of anhydrous KH₂PO₄ and 4.355 g of anhydrous K₂HPO₄ in 100 ml of distilled water. Combine the solutions and dilute to 1 L with distilled water