Recombinant Human Insulysin/IDE Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Citations (7)
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Recombinant Human Insulysin/IDE Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
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.
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
Predicted Molecular Mass
114 kDa
105 kDa, reducing conditions

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij-35 and Glycerol.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

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
Reconstitution Calculator

Reconstitution Calculator

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Background: Insulysin/IDE

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.
Long Name
Insulin Degrading Enzyme
Entrez Gene IDs
3416 (Human); 15925 (Mouse); 25700 (Rat)
Alternate Names
Abeta-degrading protease; EC 3.4.24; EC; FLJ35968; IDE; INSDEGM; Insulin protease; Insulinase; insulin-degrading enzyme; Insulysin

Citations for Recombinant Human Insulysin/IDE Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. Medium-Chain Length Fatty Acids Enhance Abeta Degradation by Affecting Insulin-Degrading Enzyme
    Authors: J Mett, AA Lauer, D Janitschke, LV Griebsch, EL Theiss, HS Grimm, H Koivisto, H Tanila, T Hartmann, MOW Grimm
    Cells, 2021;10(11):.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  2. Discovery of O-glycans on atrial natriuretic peptide (ANP) that affect both its proteolytic degradation and potency at its cognate receptor
    Authors: 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
    J. Biol. Chem., 2019;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. The insulin-degrading enzyme is an allosteric modulator of the 20S proteasome and a potential competitor of the 19S
    Authors: 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
    Cell. Mol. Life Sci., 2018;0(0):.
    Species: Rat
    Sample Types: Proteasomes
    Applications: Bioassay
  4. Ex vivo (18)O-labeling mass spectrometry identifies a peripheral amyloid ? clearance pathway
    Authors: E Portelius, N Mattsson, J Pannee, H Zetterberg, M Gisslén, H Vanderstic, E Gkanatsiou, GA Crespi, MW Parker, LA Miles, J Gobom, K Blennow
    Mol Neurodegener, 2017;12(1):18.
    Species: Human
    Sample Types: CSF
    Applications: Enzyme Assay
  5. HIV-1 reduces Abeta-degrading enzymatic activities in primary human mononuclear phagocytes.
    Authors: Lan X, Xu J, Kiyota T, Peng H, Zheng JC, Ikezu T
    J. Immunol., 2011;186(12):6925-32.
    Species: Virus
    Sample Types: Protein
    Applications: Enzyme Assay
  6. Immunocapture-based fluorometric assay for the measurement of neprilysin-specific enzyme activity in brain tissue homogenates and cerebrospinal fluid.
    Authors: Miners JS, Verbeek MM, Rikkert MO, Kehoe PG, Love S
    J. Neurosci. Methods, 2008;167(2):229-36.
    Species: Human
    Sample Types: Peptide
    Applications: Enzyme Assay
  7. Characterization of insulin degrading enzyme and other amyloid-beta degrading proteases in human serum: a role in Alzheimer's disease?
    Authors: Liu Z, Zhu H, Fang G, Walsh K, Mwamburi M, Wolozin B, Abdul-Hay S, Ikezu T, Leissring M, Qiu W
    Oncogene, 0;29(2):329-40.
    Species: Human
    Sample Types: Serum
    Applications: Control


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