Bacillus anthracis, Protective Antigen (PA 63) (FITC)

Catalog No : USB-215908
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Product name Bacillus anthracis, Protective Antigen (PA 63) (FITC)
Catalog No USB-215908
Supplier’s Catalog No 215908
Supplier US Biologicals
Source antigen
Reactivity
Cross reactivity
Applications
Molecular weight 63
Storage -20°C
Other names
Grade
Purity Single major band when examined on 4-15% polyacrylamide gels in the presence of SDS
Form Supplied as a lyophilized powder in 10mM bis-Tris propane, pH 8.5, 1.25% trehalose. Reconstitute with 200ul sterile ddH2O. Labeled with fluorescein isothiocyanate (FITC). Handle Gently. Do not vortex.
Reactivity life 12 months
Note For reserch purpose only
Purity Single major band when examined on 4-15% polyacrylamide gels in the presence of SDS
Description 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: Bacillus anthracis, Protective Antigen 63 Molecular Weight: ~63kD Binding Ratio: Contains 21ug of FITC bound/mg of PA 63 fragment of anthrax protective antigen, assuming an extinction coefficient at 493nm of 85,200M-1 cm-1 for FITC. Applications: Suitable for use in tissue culture applications. Other applications not tested. Recommended Dilution: For tissue culture applications medium containing glutamine must be fresh. Ammonium ion released when glutamine breaks down may prevent acidification of the endosome thereby inhibiting translocation of LF or EF into the cytosol. A stable form of glutamine may be used. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder is stable for 12 months after receipt at -20°C. Reconstitute with sterile ddH2O prior to use. To avoid repeated freezing and thawing, aliquot reconstituted stock solution and store at -20°C. Reconstituted product is stable at -20°C for 12 months. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: FITC conjugates are sensitive to light and should be kept in the dark. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.