Human GAK knockout HEK-293T cell line
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GAK KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Auxilin 2, Cyclin-G-associated kinase, DNAJ26, DNAJC26, EC 2.7.11.1, FLJ16629, FLJ40395, GAK protein, GAK_HUMAN, MGC99654
- Sanger seq
Unknown
Sanger Sequencing - Human GAK knockout HEK-293T cell line (AB266417)
Homozygous : Insertion of the selection cassette in exon2
- WB
Supplier Data
Western blot - Human GAK knockout HEK-293T cell line (AB266417)
This data was developed using ab288566, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration : 5% NFDM/TBST
All lanes:
Western blot - Anti-GAK antibody [EPR25327-102] (<a href='/en-us/products/primary-antibodies/gak-antibody-epr25327-102-ab288566'>ab288566</a>) at 1/1000 dilution
Lane 1:
Western blot - Human wild-type HEK-293T cell line (ab255449) at 20 µg
Lane 2:
Western blot - Human GAK knockout HEK-293T cell line (ab266417) at 20 µg
Lane 3:
Daudi (Human Burkitt’s lymphoma lymphoblast), whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Predicted band size: 143 kDa
Observed band size: 160 kDa
false
Exposure time: 59s
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com