|Product Overview:||Recombinant Anthrax Edema Factor (S414N) was expressed in B. anthracis, which has reduced adenylate cyclase activity and has been shown to act as an adjuvant.|
|Description:||Edema factor (EF) is a calmodulin-dependent adenylate cyclase which can create excess amounts of adenylate cyclase within susceptible cells.|
|Form:||When reconstituted with 0.5 ml of sterile purified water, the concentration of buffer is 5 mM HEPES, 50 mM NaCl, pH 7.5.|
|Bio-activity:||The enzymatic activity of edema factor is assessed in a cell free system by measuring cAMP formation from ATP. The activity of this EF lot is 227 uM cAMP/min/mg protein. The potency of EF to raise cAMP levels in Chinese hamster ovary (CHO) cells is tested in the presence of 1 ug/ml PA by the method of Kumar, et al. The effective concentration 50% (EC50) of this EF lot is 10.1 uM cAMP/mg CHO cell protein/mg EF protein.|
|Molecular Mass:||89 kDa|
|Endotoxin:||The endotoxin content, determined using a kinetic chromagenic LAL assay, is 153.5 EU/mg.|
|Applications:||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.|
|Storage:||This product is packaged aseptically, lyophilized and sealed under vacuum. Store at 2-8 centigrade prior to reconstitution.|
|Concentration:||Protein concentration was determined by absorbance at 280 nm using an extinction coefficient of 0.874 for 1 mg/mL.|
|Reconstitution:||Anthrax toxin proteins may be reconstituted in sterile purified 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 enzymatic activity of EF. If it is necessary to store this material, reconstitute it at a concentration of 1 mg/ml. Reconstitution with 1 mg/ml BSA will enhance stability and recovery.
It is further recommended that the solution is aliquoted and frozen at either -20 centigrade or -70 centigrade.
Avoid repeated freeze-thaw cycles. After the protein has been reconstituted as described above, glycerol may be added to 50% if a liquld is desired at freezer temperatures.
|Warning:||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.
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