Human MAP4K3 (GLK) knockout HeLa cell line
- Advanced Validation
- What is this?
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MAP4K3 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 1. 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
Germinal center kinase like kinase, Germinal center kinase-related protein kinase, MAP4K3, MAPK/ERK kinase kinase kinase 3, MAPKKKK3, MEK kinase kinase 3, MEKKK 3, RAB8IPL1, mitogen-activated protein kinase kinase kinase kinase 3
- WB
Lab
Western blot - Human MAP4K3 (GLK) knockout HeLa cell line (AB264796)
Lane 1 : Wild-type HeLa cell lysate (20ug)
Lane 2 : MAP4K3 knockout HeLa cell lysate (20ug)
Lane 3 : HepG2 cell lysate (20ug)
Lane 4 : Daudi cell lysate (20ug)
Lanes 1-2 : Merged signal (red and green). Green - MAP4K3 (D1L4G) Rabbit monoclonal antibody observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
MAP4K3 (D1L4G) Rabbit monoclonal antibody was shown to specifically react with MAP4K3 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264796 (knockout cell lysate ab258956) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and MAP4K3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. MAP4K3 (D1L4G) Rabbit monoclonal antibody and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 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:
MAP4K3 (D1L4G) Rabbit mAb at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MAP4K3 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MAP4K3 (GLK) knockout HeLa cell line (ab264796)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human MAP4K3 (GLK) knockout HeLa cell line (AB264796)
Homozygous : Insertion of the selection cassette in exon 1.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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'.
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