Recombinant Human Insulysin/IDE Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Insulysin/IDE Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >1,000 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Insulysin/IDE protein
Met42-Leu1019, with an N-terminal Met and 7-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
IDE
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
114 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
105 kDa, reducing conditions
Publications
Read Publications using
2496-ZN in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij-35 and Glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 1 M NaCl pH 7.5
  • Recombinant Human Insulysin/IDE (rhInsulysin) (Catalog # 2496-ZN)
  • Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005) , 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhInsulysin to 0.2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load 50 µL of the 0.2 µg/mL rhInsulysin into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:
  • rhInsulysin: 0.01 µg
  • Substrate: 10 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Insulysin/IDE Protein, CF

  • Abeta-degrading protease
  • EC 3.4.24
  • EC 3.4.24.56
  • FLJ35968
  • IDE
  • INSDEGM
  • Insulin protease
  • Insulinase
  • insulin-degrading enzyme
  • Insulysin

Background

Insulysin, or insulin-degrading enzyme (IDE), is a zinc metallopeptidase of the inverzincin family. IDE is primarily located in the cytosol, but has been detected as a secreted enzyme and associated with the plasma membrane as well (1). The enzyme is expressed in many tissues, with the highest levels in liver, kidney, brain, and testis (2). IDE hydrolyzes a variety of regulatory peptides, including insulin, glucagon, atrial natriuretic factor, and transforming growth factor-alpha in vitro (1). In addition, IDE has been shown to degrade the amyloid beta (A beta ) peptide, which polymerizes into the plaques associated with Alzheimer's disease (3). Deficiencies in IDE activity may contribute to the pathogenesis of type 2 diabetes mellitus (DM2) and Alzheimer's disease. The IDE region of human chromosome 10q has been genetically linked to DM2 (4). When the IDE gene was specifically disrupted in mice, IDE -/- animals developed hyperinsulinemia and glucose intolerance, characteristics of DM2 (5). The IDE -/- mice were also shown to have a significant decrease in A beta degradation in the brain, resulting in increased cerebral accumulation of A beta peptide. This in vivo evidence is consistent with the hypotheses that IDE is important for the degradation of insulin in cells and for the clearance of A beta peptide in the brain.

  1. Affholter, J. A. et al. (1988) Science 242:1415.
  2. Duckworth, W.C. et al. (1998) Endocr. Rev. 19:608.
  3. Akiyama, H. et al. (1990) Biochem. Biophys. Res. Commun. 170:1325.
  4. Selkoe, D.J. (2001) Neuron 32:177.
  5. Ghosh, S. et al. (2000) Am. J. Hum. Genet. 67:1174.
  6. Farris, W. et al. (2003) Proc. Natl. Acad. Sci. USA 100:4162.

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Publications for Insulysin/IDE (2496-ZN)(7)

We have publications tested in 3 confirmed species: Human, Rat, Virus.

We have publications tested in 3 applications: Bioassay, Control, Enzyme Assay.


Filter By Application
Bioassay
(3)
Control
(1)
Enzyme Assay
(3)
All Applications
Filter By Species
Human
(5)
Rat
(1)
Virus
(1)
All Species
Showing Publications 1 - 7 of 7.
Publications using 2496-ZN Applications Species
J Mett, AA Lauer, D Janitschke, LV Griebsch, EL Theiss, HS Grimm, H Koivisto, H Tanila, T Hartmann, MOW Grimm Medium-Chain Length Fatty Acids Enhance Abeta Degradation by Affecting Insulin-Degrading Enzyme Cells, 2021;10(11):. 2021 [PMID: 34831163] (Bioassay, Human) Bioassay Human
LH Hansen, TD Madsen, CK Goth, H Clausen, Y Chen, N Dzhoyashvi, SR Iyer, SJ Sangaralin, JC Burnett, JF Rehfeld, SY Vakhrushev, KT Schjoldage, JP Goetze Discovery of O-glycans on atrial natriuretic peptide (ANP) that affect both its proteolytic degradation and potency at its cognate receptor J. Biol. Chem., 2019;0(0):. 2019 [PMID: 31186350] (Bioassay, Human) Bioassay Human
D Sbardella, GR Tundo, A Coletta, J Marcoux, EI Koufogeorg, C Ciaccio, AM Santoro, D Milardi, G Grasso, P Cozza, MP Bousquet-D, S Marini, M Coletta The insulin-degrading enzyme is an allosteric modulator of the 20S proteasome and a potential competitor of the 19S Cell. Mol. Life Sci., 2018;0(0):. 2018 [PMID: 29594388] (Bioassay, Rat) Bioassay Rat
E Portelius, N Mattsson, J Pannee, H Zetterberg, M Gisslén, H Vanderstic, E Gkanatsiou, GA Crespi, MW Parker, LA Miles, J Gobom, K Blennow Ex vivo (18)O-labeling mass spectrometry identifies a peripheral amyloid ? clearance pathway Mol Neurodegener, 2017;12(1):18. 2017 [PMID: 28219449] (Enzyme Assay, Human) Enzyme Assay Human
Liu Z, Zhu H, Fang G, Walsh K, Mwamburi M, Wolozin B, Abdul-Hay S, Ikezu T, Leissring M, Qiu W Characterization of insulin degrading enzyme and other amyloid-beta degrading proteases in human serum: a role in Alzheimer&#039;s disease? Oncogene, 0;29(2):329-40. 0 [PMID: 22232014] (Control, Human) Control Human
Lan X, Xu J, Kiyota T, Peng H, Zheng JC, Ikezu T HIV-1 reduces Abeta-degrading enzymatic activities in primary human mononuclear phagocytes. J. Immunol., 2011;186(12):6925-32. 2011 [PMID: 21551363] (Enzyme Assay, Virus) Enzyme Assay Virus
Miners JS, Verbeek MM, Rikkert MO, Kehoe PG, Love S Immunocapture-based fluorometric assay for the measurement of neprilysin-specific enzyme activity in brain tissue homogenates and cerebrospinal fluid. J. Neurosci. Methods, 2008;167(2):229-36. 2008 [PMID: 17904641] (Enzyme Assay, Human) Enzyme Assay Human

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Bioinformatics

Gene Symbol IDE
Uniprot