Rabbit anti-Human Insulysin Polyclonal Antibody

Anti-Insulysin (Insulinase, Insulin-degrading Enzyme (IDE)), EC 3.4.24.56) Mid-molecule Antibody

Cat. Num. AAA4157565

• Reactivity: Human
• Applications: Western Blot
• Purity: Immunogen Affinity Purified
• Synonyms: Insulysin; Polyclonal Antibody; Anti-Insulysin (Insulinase; Insulin-degrading Enzyme (IDE)); EC 3.4.24.56) Mid-molecule Antibody; anti-Insulysin antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: The antibody recognises the middle region of IDE.
• Purity/Purification: Immunogen Affinity Purified
• Form/Format: Liquid; PBS, pH 7.4 with 0.05% sodium azide.
• Concentration: 1ug/ul (varies by lot)
• Sequence Length: 143

• Applicable Applications for anti-Insulysin antibody: Western Blot (WB)
• Application Notes: WB: 1:1000 (when using colorimetric substrates such as BCIP/NBT) and 1:5000 (for chemiluminescent substrates). Higher concentrations of antibody may be needed for samples from more distantly related species. Predicted molecular weight: 118 kDa. Dilution optimised using Chromogenic detection.
• Immunogen: Synthetic peptide based on the middle region of human Insulin degrading enzyme/IDE.
• Cellular Localization: Cytoplasmic
• Preparation and Storage: This product is stable for several weeks at 4 degree C as an undiluted liquid. Dilute only prior to immediate use.; For extended storage, aliquot contents and freeze at-20 degree C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of receipt.

Testing Data

Related Product Information for anti-Insulysin antibody

Description: The antibody is a polyclonal antibody made to the metalloproteinase Insulysin. The antibody is made to a synthetic peptide based on the middle region of human Insulysin. The antibody has been peptide-affinity purified, concentrated to 1 mg/ml, with the addition of 0.05% Sodium Azide as preservative, and 50% glycerol as cryoprotectant. Insulysin was identified nearly a century ago as an enzyme responsible for the degradation of insulin in cells, although the precise interactions between insulin and insulysin remain elusive. Human insulysin was cloned in 1988, and shown to be a 110 kDa protein that exists primarily as a homodimer, and perhaps also complexed with other molecules. The sequence is well conserved between humans, rats and mice, and the antibody recognizes these species. Insulysin is a metalloproteinase of the clan ME, family M16, which contains an active site HxxEH, a reversal of the canonical HExxH zinc binding motif. Considered a zinc metalloproteinase, the activity of insulysin can be blocked with EDTA or 1-10 phenanthroline. In addition to the active metalloproteinase domain, insulysin contains a second metalloproteinase site which is considered catalytically inactive, and is thought to assist in substrate binding. Insulysin is most closely related to the bacterial proteinase pitrilysin, (the human orthologue of which appears to be MPRP-1) and the mammalian proteinsae nardilysin. Generally thought to be a cytoplasmic protein, insulysin has been isolated from many different tissues and cell lines, and can degrade intact insulin, insulin B-chain, glucagon, denatured hemoglobin, alpha-amyloid protein, TGF-alpha and amylin. Recent work implicates insulysin in clearing beta amyloid plaques from the brain, and has generated much interest in Alzheimer's disease research. The pH optimum for insulysin is basic, pH 8.5, which also distinguishes it from other metalloproteinases. A recommended starting concentration for Western blots is 1:1000 when using colorimetric substrates such as BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples from more distantly related species.

Relevance: Insulysin was identified nearly a century ago as an enzyme responsible for the degradation of insulin in cells, although the precise interactions between insulin and insulysin remain elusive. Human insulysin was cloned in 1988, and shown to be a 118 kDa protein that exists primarily as a homodimer, and perhaps also complexed with other molecules. The sequence is well conserved between humans, rats and mice, and the antibody recognizes these species. Insulysin is a metalloproteinase of the clan ME, family M16, which contains an active site HxxEH, a reversal of the canonical HExxH zinc binding motif. Considered a zinc metalloproteinase, the activity of insulysin can be blocked with EDTA or 1-10 phenanthroline. In addition to the active metalloproteinase domain, insulysin contains a second metalloproteinase site which is considered catalytically inactive, and is thought to assist in substrate binding. Insulysin is most closely related to the bacterial proteinase pitrilysin, (the human orthologue of which appears to be MPRP1) and the mammalian proteinsae nardilysin. Generally thought to be a cytoplasmic protein, insulysin has been isolated from many different tissues and cell lines, and can degrade intact insulin, insulin B-chain, glucagon, denatured hemoglobin, alpha-amyloid protein, TGF-alpha and amylin. Recent work implicates insulysin in clearing beta amyloid plaques from the brain, and has generated much interest in Alzheimer's disease research. The pH optimum for insulysin is basic, pH 8.5, which also distinguishes it from other metalloproteinases. Insulin-degrading enzyme (IDE) has a preferential affinity for insulin such that the presence of insulin will inhibit IDE-mediated degradation of other substrates. IDE degrades a variety of other peptides including atrial natriuretic peptide and amylin. IDE catabolizes A-beta and has been implicated as a candidate enzyme responsible for the degradation and clearance of A-beta in the brain. IDE has also been shown to degrade the APP intracellular domain (AICD), a product of gamma-secretase cleaved APP that may function in nuclear signaling.

NCBI and Uniprot Product Information

• NCBI GI #: 358342172
• NCBI Official Full Name: insulysin