Recombinant Anthrax Protective antigen was expressed in B. anthracis. This types of PA are produced as Quality Documented (QD) reagents, distingulshed by greater control of components, equlpment and environment during production and added quality control testing for cellular activity of the final products.
Protective antigen (PA) is the receptor binding component of anthrax toxin. The original 83 kDa protein is cleaved by cellular furin-like proteases to form an activated 63 kDa monomer, PA63. In this form, PA63 binds to other PA63 molecules forming a ring-shaped heptamer. PA63 also has binding sites specific for the enzymes LF and EF adjacent to the pore formed on the cell surface for translocation. PA should be used in approximately seven-fold excess along with either EF or LF to intoxicate cells.
When reconstituted with 0.5 ml of sterile water, each vial contains 0.6mg of protein in 5 mM HEPES, 50 mM NaCl, pH 7.5.
The endotoxin content, determined using a kinetic chromagenic LAL assay, is <1 EU/mg.
Tissue Culture Application: For tissue culture applications, medium containing glutamine must be fresh. Ammonium ion is released when glutamine breaks down, and may prevent acidification of the endosome thereby inhibiting translocation of LF or EF into the cytosol. A stable form of glutamine may be used.
This product is packaged aseptically, lyophilized and sealed under vacuum. Store at 2-8 centigrade prior to reconstitution.
Anthrax toxin proteins may be reconstituted in sterile distilled water, stored at 2-8 centigrade and used successfully within a few hours. However, over longer periods of time, there may be a decline in the activity of the PA+lethal factor comlex in living cells. If it is necessary to store this material, reconstitute it at a concentration of 1 mg/ml. 2 Reconstitution with 1 mg/ml BSA may enhance stability and recovery. It is further recommended that the solution is aliquoted and frozen at either -20 centigrade or -80 centigrade. After the protein has been reconstituted as described above, glycerol may be added to 50% if a liquld is desired at freezer temperatures. Storage of material reconstituted in BSA at 4 centigrade for a period of two weeks may be acceptable for some applications.
Good laboratory technique should be employed in the safe handling of this product. This requlres observing the following practices:
1. Wear appropriate laboratory attire including a lab coat, gloves and safety glasses.
2. Do not mouth pipette, inhale, ingest or allow to come into contact with open wounds. Wash thoroughly any area of the body which comes into contact with the product.
3. Avoid accidental autoinoculation by exercising extreme care when handling in conjunction with any injection device.
4. This product is intended for research purposes by qualified personnel only. It is not intended for use in humans or as a diagnostic agent. Our company is not liable for any damages resulting from the misuse or handling of this product.