Product #120

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CERTIFICATE OF ANALYSIS ALPHA TOXIN from Staphylococcus aureus Lot #1208A1

Contents: Each vial, when reconstituted with 500.0 : l of sterile purified water, contains 0.25 mg of alpha toxin in 0.05 M sodium phosphate, pH 7.2. Concentration: Protein concentration is determined using the extinction coefficient of 1.1 at 280 nm and 1 cm for 1 mg/ml. 1 Preparation: This product is prepared from S. aureus by modifications of the procedures of Harshman et al. and Hohman. 1 ,2 Purity: This preparation migrates as a single major band of molecular weight approximately 33,000 daltons when run on 12% polyacrylamide SDS gels prepared according to the method of W yckoff, 3 a modification of the Laemmli4 gel system. When conditions for sample preparation for these gels do not exceed 60C, a faint band containing hexamer may be apparent. This preparation migrates as a single major band in slab gel electrophoresis under nondenaturing conditions. Activity: Each lot of alpha toxin is tested by hemolysis using rabbit red blood cells to confirm biological activity. 5 ,6 The specific activity of this lot is 26,000 hemolytic units per milligram. Packaging and Storage: This preparation is filter sterilized (0.2 : filter) and packaged under aseptic conditions. It is supplied as a lyophilized powder, sealed under vacuum . Store at 4C prior to and follow ing reconstitution. Maintain sterility. Prolonged storage in liquid form results in polymerization and consequent inactivation of this toxin. Toxicity: ALPHA TOXIN IS A VERY POTENT TOXIN. Alpha toxin is one of the pathogenic factors produced by S. aureus and is believed to be lethal for humans. In animals it causes respiratory paralysis, vascular and smooth muscle spasms, and tissue necrosis. Many cell types are sensitive to lysis by alpha toxin. The 50% lethal dose intravenously in mice is approximately 50 : g/kg. 7 (continued)

Handling: Good laboratory technique should be employed in the safe handling of this product. This requires observing the following practices: 1. 2. W ear appropriate laboratory attire including a lab coat, gloves and safety glasses. Do not mouth pipette, inhale, ingest or allow to come into contact with open wounds. Wash thoroughly any area of the body which com es into contact with the product. Avoid accidental autoinoculation by exercising extreme care when handling in conjunction with any injection device. This product is intended for research purposes by qualified personnel only. It is not intended for use in humans or as a diagnostic agent. List Biological Laboratories, Inc. is not liable for any dam ages resulting from the misuse or handling of this product. FOR RESEARCH PURPOSES ONLY. NOT FOR HUMAN USE. References: 1. 2. 3. 4. 5. 6. 7. Harshman, S., Sugg, N. and Cassidy, P. (1988) Meth. Enzymol. 165, 3-7. Hohman, R.J. (1988) Proc. Natl. Acad. Sci. USA 85, 1624-1628. W yckoff, M., Rodbard, D. and Chrambach, A. (1977) Anal. Biochem. 78, 459-482. Laemmli, U.K. (1970) Nature 227, 680-685. Cassidy, P. and Harshman, S. (1976) Infect. Immun. 13, 982-986. Bernheimer, A.W . (1988) Meth. Enzymol. 165, 213-217. Freer, J.H. and Arbuthnott, J.P. (1983) Pharmac. Ther. 19, 55-106.

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