Human ADAM17 knockout HCT116 cell line
- Advanced Validation
- What is this?
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(0 Publication)
ADAM17 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 5. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
A disintegrin and metalloproteinase domain 17, A disintegrin and metalloproteinase domain 17 (tumor necrosis factor, alpha, converting enzyme), ADA17_HUMAN, ADAM 17, ADAM metallopeptidase domain 17, ADAM17 protein, CD 156b, CD156b antigen, CSVP, Disintegrin and metalloproteinase domain-containing protein 17, MGC71942, NISBD, NISBD1, Snake venom-like protease, TACE, TNF-alpha convertase, TNF-alpha-converting enzyme, Tumor Necrosis Factor Alpha Converting Enzyme
- WB
Lab
Western blot - Human ADAM17 knockout HCT116 cell line (AB266873)
Lanes 1 - 2 : Merged signal (red and green). Green - ADAM17 observed at 120 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa. Rabbit anti-ADAM17 antibody was shown to react with TACE in wild-type cells in Western blot with loss of signal observed in ADAM17 knockout cell line ab266873 (ADAM17 knockout cell lysate ab256832). Wild-type and ADAM17 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with rabbit anti-ADAM17 antibody and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-tubulin-antibody-dm1a-loading-control-ab7291'>ab7291</a>) at 1/1000 dilution
Lane 1:
Wild-type cell lysate
Lane 2:
Western blot - Human ADAM17 knockout HCT116 cell line (ab266873)
Secondary
Lane 1:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lane 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 93 kDa
Observed band size: 120 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ADAM17 knockout HCT116 cell line (AB266873)
Homozygous : 1 bp deletion in exon5
Reactivity data
Product details
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
McCoY5a + 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
The ADAM17 protein regulates a multitude of cellular responses such as inflammation and growth factor signaling. It is not part of a complex but interacts with various substrates in the cell membrane. ADAM17 cleaves precursors to release soluble cytokines and growth factors like tumor necrosis factor-alpha (TNF-alpha) and epidermal growth factor receptor (EGFR) ligands which modulate cell proliferation migration and apoptosis.
Pathways
ADAM17 plays an integral role in both the TNF and EGFR signaling pathways. These pathways are important in maintaining normal cellular homeostasis. ADAM17 interacts with proteins like TNF receptors and ligands of the EGFR family integrating signaling that affects immune response and cell growth. The regulatory function of ADAM17 in these pathways makes it an important target for modulating cellular responses triggered by external stimuli.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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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