Product Information

McCoy’s 5A Medium (Iwakata and Grace Modification) with 1.5 mM L-glutamine, without sodium bicarbonate CATALOG NO. 56679C

Description Formulation In 1959, McCoy published the required amino acids for the Component (all components measured in mg/L) culture of hepatoma cells. These studies were carried out in INORGANIC SALTS Basal Medium 5A, originally developed to support Walker Calcium chloride anhydrous 100.000 Magnesium sulfate anhydrous 97.670 carcinoma 256 cells. Modifications to this medium resulting Potassium chloride 400.000 from McCoy’s work led to the development of what is now Sodium chloride 6460.000 Sodium phosphate monobasic monohydrate 580.000 known as McCoy’s 5A Medium. These formulations include VITAMINS the modifications used by Iwakata and Grace for their work Ascorbic acid 0.500 Biotin 0.200 with human myeloblasts, specifically increased levels of D-calcium pantothenate 0.200 bacto-peptone, folic acid and cyanocobalamin. Choline chloride 5.000 Cyanocobalamin 2.000 Folic acid 10.000 Precautions i-inositol 36.000 Niacin 0.500 Use aseptic technique when handling or supplementing this Niacinamide 0.500 medium after filtration. This product is for further PABA 1.000 Pyridoxal HCl 0.500 manufacturing use. THIS PRODUCT IS NOT INTENDED FOR Pyridoxine HCl 0.500 HUMAN OR THERAPEUTIC USE. Riboflavin 0.200 Thiamine HCl 0.200 AMINO ACIDS Storage L-alanine 13.360 L-arginine HCl 42.140 Store dry powder medium at 2 to 8 C. Do not use after L-asparagine monohydrate 45.030 L-aspartic acid 19.970 expiration date. Store hydrated medium protected from light L-cysteine HCl monohydrate 35.140 at 2 to 8 C. L-glutamic acid 22.070 L-glutamine 219.150 Glycine 7.510 Indications of Deterioration L-histidine HCl monohydrate 20.960 Hydroxy L-proline 19.670 Dry powder medium should be free flowing. Do not use if L-isoleucine 39.360 powder is caked. Prepared medium should be clear of L-leucine 39.360 L-lysine HCl 36.540 particulates and flocculent material. Do not use if liquid L-methionine 14.920 medium is cloudy or contains precipitate. Other evidence of L-phenylalanine 16.520 L-proline 17.270 deterioration may include color change or degradation of L-serine 26.280 physical or performance characteristics. L-threonine 17.870 L-tryptophan 3.060 L-tyrosine 2Na dihydrate 26.100 L-valine 17.570 OTHER Bacto-peptone 600.000 Dextrose anhydrous 3000.000 Glutathione reduced 0.500 Phenol red sodium salt 10.620 ADD: Sodium bicarbonate 2200.000 Grams of powder per liter 12.006

United States Europe Asia Pacific SAFC Biosciences, Inc. SAFC Biosciences Ltd. SAFC Biosciences Pty. Ltd. 13804 W. 107th Street Smeaton Road, West Portway 18-20 Export Drive Lenexa, Kansas 66215 Andover, Hampshire SP10 3LF Brooklyn, Victoria 3025 USA UNITED KINGDOM AUSTRALIA Phone +1 913-469-5580 Phone +44 (0)1264-333311 Phone +61 (0)3-9362-4500 www.safcbiosciences.com Toll free-USA 1 800-255-6032 Fax +44 (0)1264-332412 Toll free-AUS 1 800-200-404 Fax +1 913-469-5584 E-mail [email protected] Fax +61 (0)3-9315-1656 E-mail [email protected] E-mail [email protected] Preparation Instructions Characteristics Appearance 1. Measure 80 - 90% of the final volume of cell culture grade Off-white free-flowing powder water (Catalog No. 59900C) into an appropriate size mixing Bioburden vessel. Water temperature should be 15 to 30 C. Do not ≤ 100 CFU/100 mL heat water. Endotoxin 2. Add the dry powder medium to the water. Rinse the original ≤ 1.0 EU/mL package with a small amount of cell culture grade water Osmolality (as supplied) to remove all traces of powder and add to the solution. Mix Refer to Certificate of Analysis for lot-specific value until completely dissolved. pH (as supplied) 3. For each liter being prepared, add 2.2 g/L of sodium 4.5 - 4.9 bicarbonate (Catalog No. 90421C) or 29.3 mL/L of sodium bicarbonate solution 7.5% (Catalog No. 59221C). Mix References until completely dissolved. 1. McCoy, T. A., Maxwell, M., and Kruse, P. F., PSEBM (1959) 4. While stirring the solution in Step 3, adjust the pH to 100:115. 6.7 - 6.9 using NaOH 1N (Catalog No. 59223C) or 2. Neuman, R. E. and McCoy, T. A., PSEBM (1958) 98:303. HCl 1N. The pH of bicarbonate buffered solutions usually 3. Hsu, T. C. and Kellogg, D. S., JNCI (1960) 25:221. rises 0.1 - 0.2 units during filtration. 4. Iwakata, S. and Grace, J. J., NY State J Med (1964) 64:2279 5. Add cell culture grade water to the solution in Step 4 to bring it to the final volume. Keep the vessel closed until the solution is filtered to avoid fluctuations in pH. 6. Sterilize the solution using a membrane filter with a pore size of 0.22 µm or less. A peristaltic pump or an inert gas such as nitrogen can be used to provide positive pressure at 3 - 15 psi. Do not use CO2 gas. 7. Sterile solutions should be dispensed aseptically into sterile containers. Store protected from light at 2 to 8 C. 8. Supplements, such as antibiotics, can be added to the sterilized solution using aseptic technique. Storage conditions and shelf life of the supplemented product may be affected by the nature of the supplements. Sterile serum should not be refiltered before or after being added to sterile medium because growth promoting capacity may be reduced upon refiltration. NOTE: Dry powder medium is extremely hygroscopic and must be protected from atmospheric moisture. We recommend that the entire contents of each package be used immediately after opening.

Warranty, Limitation of Remedies SAFC Biosciences warrants to the purchaser for a period of one year from date of delivery that this product conforms to its specifications. Other terms and conditions of this warranty are contained in SAFC Biosciences’ written warranty, a copy of which is available upon request. ALL OTHER WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING THE IMPLIED WARRANTY OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE, ARE EXCLUDED. In no case will SAFC Biosciences be liable for any special, incidental, or consequential damages arising out of this product or the use of this product by the customer or any third party based upon breach of warranty, breach of contract, negligence, strict tort, or any other legal theory. SAFC Biosciences expressly disclaims any warranty against claims by any third party by way of infringement or the like. THIS PRODUCT IS INTENDED FOR PURPOSES DESCRIBED ONLY AND IS NOT INTENDED FOR ANY HUMAN OR THERAPEUTIC USE. Additional Terms and Conditions are contained in the product Catalog, a copy of which is available upon request.

Issued December 2006 P56679 1203 1005 0406 0606 0906