Dialysis of serum

Summary

This lab was derived from: Animal Cell Culture - A Guide to Basic Techniques (5th Edition)

Operation method

Scheme 11.16 Dialysis of serum

Materials and Instruments

Serum HBSS
Bottles Lids Sterile filters Dialysis bags Large beakers Bunsen burners Tripods Measuring cylinders

Move

1. Boil five 30 mm × 500 mm dialysis bags and replace with distilled water three times.

2. Transfer the dialysis bags to Hanks' balanced salt solution (HBSS) (HBSS stored at 4°C) and cool.

3. tie two knots at one end of each dialysis bag.

4. fill each dialysis bag halfway with serum (serum to be dialyzed) (20 ml ).

5. Press in the air and then tie the other end of the dialysis bag in a knot as well, leaving about half the space of the tubing between the serum and the knot.

6. Place the dialysis bag in 5 L of HBSS and mix on a magnetic stirrer at 4°C overnight.

7. Replace the HBSS and repeat step 6 twice.

8. Collect the serum into a measuring cylinder and record the amount collected (if it is less than the amount of serum at the beginning, add HBSS to bring it back to the beginning amount. If it is more than the amount of serum at the start, leave a reserve for subsequent additions to the medium ).

9. Sterilize the serum using a series of filters (sterile filters, 0.1 μm pore size; prefilters: glass fibers with a pore size of 5.0 μm, and membranes (e.g., Durapore, Millipore) of 1.2 μm, 0.45 μm, and 0.22 μm).

10. Serum was bottled and frozen.

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