PREPARATION OF HB SUPPLEMENT

LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

Appendix

REFERENCE NO: AHC/1998/3/1.2 Appendix 6

TITLE: PREPARATION OF HB SUPPLEMENT

INTRODUCTION

This media supplement may be added to tissue culture media for slow growing cell lines such as hybridomas.

PROCEDURE

Materials

1320 mg oxaloacetic acid
80 mg bovine insulin
550 mg sodium pyruvate
100 labelled and dated 1 ml cryovials
0.2 mm filters, 20 ml syringes

Add the oxaloacetic acid and bovine insulin to 80 ml of distilled water.
Dissolve in a 37°C water bath at between 35 to 38°C.
Add 550 mg of sodium pyruvate.
Adjust to 100 ml final volume with dH₂O.
Filter sterilise, aliquot and store at <-10°C.

The solution is used at 1:100 final dilution in media. Mix well on thawing as precipitates may form and add to tissue culture medium.

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