Principles of Biochemistry (2+1)

Reagents

1. CM-cellulose-5ml

2. 1M Tris-HCl, pH 9. buffer: Combine 40ml of 5M NaCl, 50ml 1M Tris(pH9), and 0.7ml beta-mercaptoethanol. Bring volume to 1L with distilled water. Store in a tightly capped bottle in refrigerator. 

3. 1M stock phosphate solutions: Prepare 100ml of 1M Potassium dibasic and (K₂HPO₄) monobasic (KH₂PO₄)solutions separately. Prepare 0.1M working stocks of the dibasic and monobasic solutions by diluting the 1 M stocks of  (K₂HPO₄) and  (KH₂PO₄) solutions

4. 0.1M elution buffer pH 7: Mix the two diluted working stocks in a ratio of 61.5 ml 0f K₂HPO4 and 38.5 ml of KH₂PO₄ Check the pH of the solution. Continue adding each solution, as appropriate, and checking the pH until pH 7 is reached.

5. Neutralizing solution

6. 5M sodium chloride

7. 0.5M phosphate buffer : Prepare by diluting 1M phosphate buffer.

8. Lysozyme standard