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 |