Anti-Clostridium botulinum Toxin D Antibody

[Botulinum neurotoxin type D]: Botulinum toxin causes flaccid paralysis by inhibiting neurotransmitter (acetylcholine) release from the presynaptic membranes of nerve terminals of the eukaryotic host skeletal and autonomic nervous system, with frequent heart or respiratory failure (PubMed:8175689, PubMed:16252491). Precursor of botulinum neurotoxin D for which a proteinaceous coreceptor is controversial. In double SV2A/SV2B knockout mice this toxin does not degrade its synaptobrevin target; introducing SV2A, SV2B or SV2C restores target cleavage (PubMed:21483489). RUniProtKB:P0DPI0, PubMed:16252491, PubMed:19650874, PubMed:20704566, PubMed:21483489, PubMed:21632541, PubMed:8175689, -Leu-62' bond of synaptobrevin-1 (VAMP1), and the equivalent 'Lys-|-Leu' sites in VAMP2 and VAMP3 (PubMed:8175689). Cleaves the '49-Lys-|-Ile-50' bond of A.californica synaptobrevin (AC P35589) (PubMed:8197120). This chain probably has to be partially unfolded to translocate into the eukaryotic host cell cytosol (PubMed:15584922). {PubMed:8175689, PubMed:8197120, PubMed:15584922}.; [Botulinum neurotoxin D heavy chain]: Responsible for host epithelial cell transcytosis, host nerve cell targeting and translocation of light chain (LC) into eukaryotic host cell cytosol. Composed of 3 subdomains; the translocation domain (TD), and N-terminus and C-terminus of the receptor-binding domain (RBD). The RBD is responsible for the adherence of the toxin to the eukaryotic target cell surface. The N-terminus of the TD wraps an extended belt around the perimeter of the LC, protecting Zn(2+) in the active site; it may also prevent premature LC dissociation from the translocation channel and protect toxin prior to translocation (PubMed:17907800). The TD inserts into synaptic vesicle membrane to allow translocation into the host cytosol (By similarity). The RBD binds eukaryotic host phosphatidylethanolamine, which may serve as toxin receptor (PubMed:16115873). Treatment of synaptosomes with proteinase K does not reduce HC binding, suggesting there is no protein receptor or it is protected from extracellular proteases (PubMed:16115873). HC significantly decreases uptake and toxicity of whole BoNT/D (PubMed:19650874, PubMed:21483489). HC also interferes with uptake of tetanus toxin (PubMed:19650874). Has 2 closely located carbohydrate-binding receptor sites and binds at least 1 GT1b ganglioside (PubMed:20704566). Bind gangliosides in the order GD2 > GT1b > GD1b (PubMed:21632541). Interacts with eukaryotic target protein SV2B (synaptic vesicle glycoprotein 2B) (PubMed:21483489). Expression of SV2A, SV2B or SV2C in mice knocked-out for the SV2 proteins restores entry of BoNT/D and cleavage of VAMP2, suggesting SV2 acts as its receptor (PubMed:21483489). Unlike BoNT/A and BoNT/E, toxin uptake is not mediated by large extracellular loop 4 of SV2 (PubMed:21483489). Another group finds very poor interaction with SV2 proteins, suggesting the possible protein receptor may not have been identified (PubMed:21632541). {UniProtKB:P0DPI0, PubMed:16115873, PubMed:19650874, PubMed:20704566, PubMed:21483489, PubMed:21632541, PubMed:17907800}.

ELISA: use a dilution of 1:20-1:200. Immunocytochemistry/ <br><br>Immunohistochemistry: use a dilution of 1:10-1:50. <br><br>Immunofluorescence: use a dilution of 1:10-1:50 <br><br>Immunoblotting: use a dilution of 1:100-1:1,000. A band of ~150,000 is detected. <br><br>End users should determine optimal dilutions for their applications.

This antibody is stable at 4°C for short-term storage and for at least one (1) year at -20° to -70°C. Store product in appropriate aliquots to avoid multiple freeze-thaw cycles.

Clostridium botulinum, an anaerobic, gram-positive, spore-forming rod commonly found on plants, in soil, water, and the intestinal tracts of animals, produces eight antigenically distinguishable exotoxins (A, B, C1, C2, D, E, F and G). Type A is the most potent toxin, followed by types B and F. All botulinum neurotoxins are produced as single polypeptide chains of ~150kDa comprised of a heavy (H) chain and a light (L) chain of roughly 100 and 50kDa, respectively, linked by a disulfide bond. The heavy (H) chain of the toxin binds selectively and irreversibly to high affinity receptors at the presynaptic surface of cholinergic neurones, and the toxin-receptor complex is taken up into the cell by endocytosis where the disulfide bond between the two chains is cleaved. The light (L) chain interacts with different proteins in the nerve terminals to prevent fusion of acetylcholine vesicles with the cell membrane.