Mouse Insulin Degrading Enzyme (IDE) ELISA Kit ( Part mIDE-Biotin) kw: insulin degrading enzyme, neurosin

Mouse Insulin Degrading Enzyme (IDE) ELISA Kit

Dynamic Assay Range: 300pg/ml - 60ng/ml. Sensitivity: 220pg/ml (July 2014)

Lead Time: Approximately 2.5 weeks

Specifications

Fully pre-configured ELISA kit in sandwich assay format.
Dynamic Assay Range: 300pg/ml - 60ng/ml. Sensitivity: 226pg/ml.
Rapid ELISA Protocol: Requires one 60 min incubation period at 37^oC.
Samples are mixed with the provided biotinylated detection antibody and provided streptavidin-HRP prior to addition to plate.

Number of Assays	Cost
96T	$625
2 x 96T	$1170

Methods Overview:

The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen. Samples are pre-mixed with a provided biotin-linked detection antibody and provided streptavidin-HRP, which are then added to the plate. The plate is washed after a 60 min incubation period. Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution. A stop solution is applied to terminate the reaction, and the solution is measured at 450nm.

Protocol: (Updated assay range and standard are posted on this webpage).

Sandwich ELISA Methods to Insulin Degrading Enzyme (IDE)

ELISA Assay Steps

Mix sample, provided biotinylated-detection antibody and provided streptavidin-HRP.
Apply sample and antibody-HRP complex to ELISA plate for 60min at 37^oC. Wash 5 times.
Apply chromogens, incubate, then stop reaction.

Introduction to Insulin Degrading Enzyme:

Insulin-degrading enzyme (IDE) is a zinc-binding protease known to cleave short polypeptides with variable sequences. IDE was first identifiable for its ability to degrade the B chain of insulin and 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 show 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 indiivduals.

Gerneral References on Insulin Degrading Enzyme

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.
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.
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.
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.
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.
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.

Safety: Stop solution contains acid. Avoid ingestion, skin and eye contact.

Storage: 4°C. 6 months.

Category	Download Link
Protocol Manual	click here
MSDS	click here

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