Human AAK1 knockout HeLa cell line
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AAK1 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 4. 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
AAK1_HUMAN, AP2 associated kinase 1, AP2-associated protein kinase 1, Adaptor-associated kinase 1, DKFZp686F03202, DKFZp686K16132, FLJ23712, FLJ25931, FLJ31060, FLJ42882, FLJ45252, KIAA1048, MGC138170, MGC164568, MGC164570
- Sanger seq
Unknown
Sanger Sequencing - Human AAK1 knockout HeLa cell line (AB265047)
Homozygous : Insertion of the selection cassette in exon 4.
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
AAK1 functions as part of the AP2 clathrin adaptor complex which is essential for the internalization of cargo at the plasma membrane. Its activity is important for maintaining proper cellular uptake of nutrients receptors and signaling molecules. In neurons AAK1 regulates the cycle of synaptic vesicles impacting neurotransmission by influencing vesicle release and recycling. It has been suggested that AAK1 also contributes to the modulation of dendritic spine morphology through its kinase activity.
Pathways
AAK1 participates in endocytosis and intracellular trafficking pathways. It is directly involved in the clathrin-mediated endocytic route interacting with other proteins such as clathrin and dynamin. This association with clathrin provides a mechanism for cargo selection and membrane curvature linking AAK1 to the broader system of intracellular transport. In addition AAK1 connects to MAPK signaling through potential cross-talk mechanisms highlighting its role in cellular signaling regulation.
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
Assay Kits
AB207212
MAPK (ATF2, c-Jun, c-Myc, MEF2, STAT1alpha) Transcription Factor Assay Kit (Colorimetric)
- 0
- 0
Primary Antibodies
AB32520
Recombinant|Advanced Validation|RabMAb|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] (AB32520)](https://content.abcam.com/products/images/stat6-antibody-ye361-ab32520--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img112052.jpg)
- 5
- 1
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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