Human IDE (Insulin degrading enzyme) knockout HeLa cell lysate
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IDE KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 14.
View Alternative Names
Abeta-degrading protease, FLJ35968, IDE_HUMAN, Insulin protease, Insulin-degrading enzyme, Insulinase, Insulysin, OTTHUMP00000020097
- WB
Lab
Western blot - Human IDE (Insulin degrading enzyme) knockout HeLa cell lysate (AB257197)
Lane 1 : Wild-type HeLa cell lysate (20μg)
Lane 2 : IDE knockout HeLa cell lysate (20μg)
Lanes 1- 2 : Merged signal (red and green). Green - ab109538 observed at 118 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109538 Anti-Insulin degrading enzyme / IDE antibody [EPR6099] was shown to specifically react with Insulin degrading enzyme / IDE in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261755 (knockout cell lysate ab257197) was used. Wild-type and Insulin degrading enzyme / IDE knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109538 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Insulin degrading enzyme / IDE antibody [EPR6099] (<a href='/en-us/products/primary-antibodies/insulin-degrading-enzyme-ide-antibody-epr6099-ab109538'>ab109538</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
IDE knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human IDE (Insulin degrading enzyme) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ide-insulin-degrading-enzyme-knockout-hela-cell-line-ab261755'>ab261755</a>)
Predicted band size: 118 kDa
Observed band size: 118 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human IDE (Insulin degrading enzyme) knockout HeLa cell lysate (AB257197)
Homozygous : 4 bp deletion in exon 14
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
IDE manages the levels of insulin and other peptides by degrading them preventing accumulation and maintaining homeostasis. It is not part of a complex but it acts individually in cellular environments to modulate the concentration of its substrates. IDE is important for controlling insulin availability and turnover which impacts glucose metabolism. By influencing the degradation of insulin IDE aids in balancing metabolic demands with insulin availability.
Pathways
IDE plays a vital role in insulin signaling and glucose metabolic processes. It is directly involved in the insulin signaling pathway by regulating insulin levels which consequently affects cellular responses to insulin. IDE connects with several proteins associated with these pathways including insulin receptor and glucose transporters ensuring proper cell signaling and metabolic functions. By modulating insulin levels IDE helps optimize glucose uptake and storage.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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