Human DAPK3 Knockout HEK293T Cell Line
|Product Size||1 × 106 cells/vial, 1 mL|
Shipping & Storage Information
|Storage Condition||Upon arrival, the vial should be stored in liquid nitrogen vapor phase.
Storage buffer: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
|Shipping Condition||Shipped on dry ice.|
|Parental Cell Line||HEK293T|
|Mutation Description||CRISPR/Cas9 system is used to generate knockout cell lines. Homozygous: 1 bp insertion in exon 1. Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest.|
|Adherent / Suspension||Adherent|
|Cell Type||Epithelial Cells|
|Knockout Validation||Sanger Sequencing|
|Cell Medium||Culture medium: DMEM (High Glucose) + 10% FBS.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
|Initial Handling Guidelines||1. Thaw the vial in 37ºC water bath approximately 1-2 minutes.
2. Transfer the cell suspension to a 15 ml/50 ml conical sterile polypropylene centrifuge tube containing pre-warmed culture medium. Wash vial with an additional culture medium to collect remaining cells, and centrifuge at 201 × g (rcf) for 5 minutes at room temperature.
3. Resuspend the cell pellet in 5 ml pre-warmed culture medium and count cell number. Seed cells in an appropriate cell culture flask at a density of 2×104 cells/cm2. This should allow for confluency within 48 hours.
4. Incubate the culture at 37ºC incubator with 5% CO2. Cultures should be monitored daily.
|Subculture Guidelines||Cells should be passaged when they have achieved 80-90% confluence. A partial media change 24 hours prior to subculture may be helpful to encourage growth.|
|Applications||Western blot. Predicted molecular weight: 53 kDa.|
|Description||Human wild-type HEK293T cell line is the recommended control.|
|Cellular Localization||Cytoplasm, nucleus, and PML body.|
|Target Sequence Similarities||Belongs to the CAMK Ser/Thr protein kinase family, DAP kinase subfamily.|
|Post-Translational Modifications||Ubiquitinated and autophosphorylated.|