Human IDE (Insulin degrading enzyme) knockout HeLa cell line
- Advanced Validation
- What is this?
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IDE KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. 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 line (AB261755)
Lanes 1- 2 : Merged signal (red and green). Green - ab109538 observed at 118 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109538 was shown to 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 HeLa and IDE knockout HeLa cell lysates 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) overnight at 4°C at a 1 in 1000 dilution and a 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 (ab261755)
Predicted band size: 118 kDa
Observed band size: 118 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human IDE (Insulin degrading enzyme) knockout HeLa cell line (AB261755)
Homozygous : 4 bp deletion in exon 14.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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
Target data
Complementary Products
Primary Antibodies
AB179483
Anti-HIF-1 alpha antibody [EPR16897]
RabMAb|Advanced Validation|Recombinant|KO Validated
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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