Bacillus anthracis is the etiological agent of anthrax. Major virulence factors produced by Bacillus anthracis are the gamma-linked, poly-D-glutamic acid capsule and an exotoxin composed of three components, protective antigen (PA), lethal factor (LF), and edema factor (EF). These proteins are not toxic separately but act in binary combinations. The complex of PA, the cell binding component, with LF, one of the enzymatic moieties, is termed “lethal toxin” and can cause death. PA and the enzymatic EF together cause skin edema. After secretion, PA is cleaved by membrane peptidases, allowing the 63kD carboxy terminal fragment to oligomerize to a heptamer. Cleavage is an essential step in exposing the binding sites for EF and LF. The assembled PA complex and associated LF or EF enters the cell through endocytosis. PA mediates the transfer of LF and EF to the cytoplasm where these enzymes recognize and proteolyze their targets. The LF component, a zinc-dependent metalloprotease, cleaves the mitogen-activated protein kinase kinase (MAPKK), impairing its function and thereby blocking the MAPK signaling pathway. EF is a calmodulin-dependent adenylate cyclase which creates in the cell excess amounts of cyclic-AMP, resulting in edema. Current research is focused on the development of an effective vaccine as well as screening for potent inhibitors of the LF enzymatic activity.
Source:Recombinant protein corresponding to Bacillus anthracis Lethal Factor, Mutant E687C.
Molecular Weight:~88kD-90kD
Applications:Suitable for use in immunodiffusion. Other applications not tested.
Recommended Dilution:Optimal dilutions to be determined by the researcher.
Biological Activity:In immunodiffusion studies, anthrax mutant lethal factor shows similar immuno-precipitation as anthrax lethal factor when reacted against a specific antiserum to anthrax lethal factor. No cytotoxicity was detected with 1ug/ml (11nM) LF mutant in the presence of 1ug/ml (12nM) protective antigen when tested on J774.1A cells. This LF mutant, E687C, has no enzymatic activity.
Protein Concentration:Determined by absorbance at 280nm using an extinction coefficient of 0.812 for 1mg/ml.
Storage and Stability:Lyophilized powder may be stored at -20°C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
仕様
Size：100ug
Source Antigen：Recombinant
Purity：When examined on 10% polyacrylamide gels in the presence of SOS, this preparation migrates as two major bands with apparent molecular weights of 88,000 and 90,000 daltons.
Form：Supplied as a lyophilized powder in 5mM HEPES, 50mM sodium chloride, pH 7.5. Reconstitute with 100ul sterile ddH2O. Handle Gently. Do not vortex.
