Human ILKAP knockout HEK-293T cell line
- Advanced Validation
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ILKAP 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: 5 bp deletion in exon 2.
View Alternative Names
DKFZp434J2031, FLJ10181, ILKAP2, ILKAP3, ILKAP_HUMAN, Integrin linked kinase associated phosphatase, Integrin linked kinase associated serine/threonine phosphatase, Integrin-linked kinase-associated serine/threonine phosphatase 2C, MGC4846, PP2C-delta, Protein phosphatase 2c delta isozyme
- WB
Lab
Western blot - Human ILKAP knockout HEK-293T cell line (AB266475)
Lanes 1-4 : Merged signal (red and green). Green - ab196013 observed at 49 kDa. Red - loading control ab8245 observed at 36 kDa.
ab196013 Anti-ILKAP antibody [EPR16145] was shown to specifically react with ILKAP in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266475 (knockout cell lysate ab258000) was used. Wild-type and ILKAP knockout samples were subjected to SDS-PAGE. ab196013 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-ILKAP antibody [EPR16145] (<a href='/en-us/products/primary-antibodies/ilkap-antibody-epr16145-ab196013'>ab196013</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ILKAP knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ILKAP knockout HEK-293T cell line (ab266475)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
HAP1 cell lysate at 20 µg
Secondary
All lanes:
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 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 49 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ILKAP knockout HEK-293T cell line (AB266475)
Homozygous : 5 bp deletion in exon 2
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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
The phosphatase activity of ILKAP plays a role in cellular adhesion and movement. ILKAP participates in cellular processes by associating with the ILK complex which includes other key proteins like paxillin and parvin. By modulating the phosphorylation state of substrates within this complex ILKAP influences focal adhesion dynamics and cytoskeletal organization which are critical for maintaining cellular architecture and motility.
Pathways
ILKAP acts within the integrin signaling and Wnt signaling pathways. These pathways are important in regulating cell movement differentiation and survival. ILKAP interacts with proteins like β-catenin in the Wnt pathway influencing gene expression critical for cell fate determination. Additionally ILKAP works alongside ILK in the integrin signaling pathway to affect cellular responses like proliferation and migration.
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
AB196013
Anti-ILKAP antibody [EPR16145]
RabMAb|Recombinant|KO Validated|Trial Size|Trial Size|Trial Size|Trial Size
![Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)](https://content.abcam.com/products/images/ilkap-antibody-epr16145-ab196013--western-blot-img5042.jpg)
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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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