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Insulin Degrading Enzyme ELISA Kit | FIVEphoton Biochemicals | IDE-ELISA, 96T, IDE

Insulin Degrading Enzyme ELISA Kit | FIVEphoton Biochemicals | IDE-ELISA, 96T, IDE
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Price: $625.00
Availability: In Stock
Model: IDE-ELISA (96T)

Available Options:
Format 1:
Format 2:
Format 3:
DetailsManuals & DownloadsRelated Products
 
Insulin Degrading Enzmye (IDE) ELISA Kits 
Highly sensitive sandwich ELISA Kits with pg/ml detection and rapid protocol.  96T

Lead time: 2 weeks.

Insulin Degrading Enzyme (IDE) ELISA Kits.  Separate kits are available for human, mouse, rat, canine and rabbit.
  • Fully pre-configured sandwich ELISA format providing optimal convenience, specificity and sensitivity
  • Strip plate configuration allowing separate use and storage of 96-well ELISA plate sections
  • Affinity purified polyclonal or monoclonal capture antibodies coated on plate, depending on kit format.  Separate HRP-conjugated detection antibody
  • Rapid colorimetric detection. 1 hr total incubation at 37oC
  • Suitable for CSF, cell and tissue lysates and homogenates, other biological fluids
  • Highly sensitive: measures Insulin Degrading Enzyme concentration in pg/ml
  • Available for Human, rat, mouse, canine and rabbit Insulin Degrading Enzyme
  Protocol IDE ELISA Format 1
 
         Protocol IDE ELISA Format 2

Specifications of Insulin Degrading Enzyme ELISA Kits

Format 1.  View this product at these links:  human:    mouse
 
 Species
 Standard 
 Assay Range
 Sensitivity
 Price
 Human 192 ng/ml 600 pg/ml -  180 ng/ml
 544pg/ml
 $720
 
Format 2
 Species
 Standard 
 Assay Range
 Price
 Human 72 pg/ml 2-50 pg/ml $655
 Rat 22.5 pg/ml 0.7-20 pg/ml $655
 Mouse 22.5 pg/ml 0.7-20 pg/ml $655
 Canine 54 pg/ml 1-45 pg/ml $655
 Rabbit
 54pg/ml
 1-45 pg/ml
 $655
 
Format 3  Please request protocol
 Species
 Standard (lyophilized Solid) 
 Assay Range
 Sensitivity
 Price
 Human 20ng: 2 vials
 62.5 pg/ml -  4 ng/ml
 <37.5 pg/ml
 $655
 Mouse 4 ng: 2 vials
 313 pg/ml - 20 ng/ml <313 pg/ml
 $655

Use Available Options menu to select species and format.

Background:  Insulin-degrading enzyme (abbreviated IDE), also known as insulysin or insulin protease, is a zinc-binding protease of the M16A metalloprotease subfamily known to cleave short polypeptides with variable sequences.  IDE was first identified by its ability to degrade the B chain of insulin.  IDE, which migrates at 110 kDa in gel electrophoresis under denaturing conditions, has since been shown to have additional substrates, including the signaling peptides glucagon, TGF alpha, β-endorphin and amyloid-beta (Aβ).

In vitro and in vivo studies have shown correlations between IDE activity levels, Aβ degradation, and Alzheimer’s disease.  Mice engineered to lack both alleles of the IDE gene exhibit a 50% decrease in Aβ degradation, resulting in cerebral accumulation of Aβ.  Studies of genetically inherited forms of Alzheimer’s show reduction in both IDE expression and catalytic activity among affected individuals.
Methods Overview 
Sandwich ELISA Format 1:  The ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen.  Samples are pre-mixed with provided  biotin-linked detection antibody and streptavidin-HRP solutions, which are then added to the plate and incubated for 60min.  The plate is washed  and color reagents are added, resulting in a reaction with HRP.  A stop solution is applied and the resulting yellow solution is measured at 450nm.

Sandwich ELISA Format 2:  The included ELISA 96-well micro-strip plate is provided pre-coated with an affinity purified rabbit polyclonal antibody selective to the antigen.  Upon addition of samples to the wells, the antigen in solution binds to the antibody and becomes indirectly linked to the solid support.  After washes of unbound materials, a second affinity purified polyclonal antibody conjugated with horse-radish peroxidase (HRP) is added, followed by a second incubation period.  Unbound antibody is washed, and then color reagents are added, which upon conversion by HRP, become blue colored.  A stop solution is added to block further reaction between HRP and the colorimetric substrates, converting the solution into a yellow coloration.  An absorbance multiplate reader is used to quantitate the colorimetric reaction at 450 nm.  This assay requires approximately 2.5 hrs to complete at RT.
 
*(Use the protocol shipped with the kit for your experiment).

Image:  Crystal structure of human insulin-degrading enzyme in complex with amyloid-beta (1-40).  PDB:  DOI:10.2210/pdb2g47/pdb
   Representative Sandwich ELISA Methods                                                                                                                           
 
Steps of Sandwich ELISA Format 1
1.  Mix sample, provided biotinylated-detection antibody and provided streptavidin-HRP.
2.  Apply sample and antibody-HRP complex to ELISA plate for 60min at 37oC.  Wash 5 times.
3.  Apply chromogens, incubate, then stop reaction.
 
 
References on Insulin Degrading Enzyme (IDE)
  1. Amata, O., Marino, T., Russo, N., & Toscano, M. (2009). Human insulin-degrading enzyme working mechanism. Journal of the American Chemical Society, 131(41), 14804-14811.
  2. Bennett, R. G., Duckworth, W. C., & Hamel, F. G. (2000). Degradation of amylin by insulin-degrading enzyme. The Journal of Biological Chemistry, 275(47), 36621-36625.
  3. Grasso, G., Rizzarelli, E., & Spoto, G. (2008). How the binding and degrading capabilities of insulin degrading enzyme are affected by ubiquitin.Biochimica et Biophysica Acta, 1784(7-8), 1122-1126.
  4. Hong, M.-G., Reynolds, C., Gatz, M., Johansson, B., Palmer, J. C., Gu, H. F., Blennow, K., et al. (2008). Evidence that the gene encoding insulin degrading enzyme influences human lifespan. Human Molecular Genetics, 17(15), 2370-2378. Oxford University Press.
  5. Im, H., Manolopoulou, M., Malito, E., Shen, Y., Zhao, J., Neant-Fery, M., Sun, C.-Y., et al. (2007). Structure of substrate-free human insulin-degrading enzyme (IDE) and biophysical analysis of ATP-induced conformational switch of IDE. The Journal of Biological Chemistry, 282(35), 25453-25463.
  6. Vepsalainen, S., Hiltunen, M., Helisalmi, S., Wang, J., Van Groen, T., Tanila, H., & Soininen, H. (2008). Increased expression of Abeta degrading enzyme IDE in the cortex of transgenic mice with Alzheimer’s disease-like neuropathology. Neuroscience Letters. 
For research only.  Not for diagnostics of disease in humans or animals.
Lead  time:  approximately 2 weeks.
Safety:  Stop solution contains acid.   Avoid ingestion, skin and eye contact.
Storage:  4oC, 6 months.  Longer term storage of sections of the strip plate and standards at -20oC.  Seal and store 96-well ELISA plate in a zip locks bag after opening.
Shipping.  Overnight domestic delivery recommended (Flat rate Fedex cost for continental US: $42).  Also available for international delivery; please inquire for shipping cost. 

KW:  Rabbit Insulin Degrading Enzyme (IDE) ELISA Kit
Human Insulin Degrading Enzyme (IDE) ELISA Kit
Mouse Insulin Degrading Enzyme (IDE) ELISA Kit
Rat Insulin Degrading Enzyme (IDE) ELISA Kit
Canine Insulin Degrading Enzyme (IDE) ELISA Kit

 

 


 

Category Download Link
Protocol Manual Format I
 click here
Protocol Manual Format 2
 click here
MSDS click here



 

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