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Active Insulin Degrading Enzyme (IDE)

Insulysin; Insulin Protease; Abeta-degrading protease; Insulinase

ACTIVITY TEST

Insulin Degrading Enzyme (IDE) is an evolutionarily conserved 110-kDa zinc metalloprotease. It has been described principally as a cytosolic enzyme but is also found in multiple cellular compartments including endosomes, peroxisomes, mitochondria, the cell surface and in secreted form. IDE is a major enzyme responsible for insulin degradation. In addition to insulin, IDE degrades many targets including glucagon, atrial natriuretic peptide, and beta-amyloid peptide, regulates proteasomal degradation and other cell functions. In addition, IDE can degrade IGF2, thus affecting the concentration and activity of IGF2 in the cell. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human IDE and recombinant rabbit IGF2. Briefly, IDE was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IGF2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IDE pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human IDE and recombinant rabbit IGF2 was shown in Figure 1, the EC50 for this effect is 0.08 ug/mL.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
J Neurochem Reduction PubMed: 26442993
African Journal of Pharmacy and Pharmacology Peptide hydroxamate derivatives as regulators for insulin receptor signaling and its degradation by zinc metalloprotease in diabetic rats article-full-text-pdf
Biomedicine & Pharmacotherapy Vitamin D3 intake as regulator of insulin degrading enzyme and insulin receptor phosphorylationin diabetic rats. pubmed:27930980
Biological Trace Element Research Nigella sativa Oil and Chromium Picolinate Ameliorate Fructose-Induced Hyperinsulinemia by Enhancing Insulin Signaling and Suppressing Insulin-Degrading Enzyme in Male Rats 10.1007/s12011-017-1167-z
Neuroscience The Protective Effects of IGF-I against β-Amyloid-related Downregulation of Hippocampal Somatostatinergic System Involve Activation of Akt and Protein Kinase A Pubmed:29406271
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KSB897Hu01 ELISA Kit DIY Materials for Insulin Degrading Enzyme (IDE) Main materials for "Do It(ELISA Kit) Yourself"