Human TLK1 knockout HeLa cell line
- Advanced Validation
- What is this?
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TLK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and Insertion of the selection cassette in exon 4.
View Alternative Names
PKU-beta, SNARE protein kinase SNAK, Serine threonine protein kinase, Serine/threonine-protein kinase tousled-like 1, TLK1_HUMAN, Tousled-like kinase 1
- WB
Lab
Western blot - Human TLK1 knockout HeLa cell line (AB264898)
Lanes 1 - 4 : Merged signal (red and green). Green - Rabbit polyclonal antibody observed at 90 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
Rabbit polyclonal antibody was shown to react with TLK1 in wild-type HeLa cells in Western blot with loss of signal observed in TLK1 knockout cell line ab264898 (TLK1 knockout cell lysate ab258231). Wild-type HeLa and TLK1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Rabbit polyclonal antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Rabbit polyclonal antibody at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
TLK1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human TLK1 knockout HeLa cell line (ab264898)
Lane 3:
HL-60 cell lysate at 20 µg
Lane 4:
PC3 cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human TLK1 knockout HeLa cell line (AB264898)
Allele-2 : Insertion of the selection cassette in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human TLK1 knockout HeLa cell line (AB264898)
Allele-1 : 1 bp insertion in exon 4.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). 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, 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
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
TLK1 contributes significantly to chromatin remodeling and DNA repair by influencing histone dynamics. TLK1 often acts in concert with other proteins as part of regulatory complexes modulating DNA accessibility during replication and repair processes. As a kinase it phosphorylates and activates targets like Asf1 a histone chaperone impacting nucleosome assembly.
Pathways
TLK1 plays a significant role in the DNA damage response and cell cycle regulation pathways. It is closely related to the ATM/ATR pathway which is central to the cellular response to DNA damage. TLK1 also interacts with proteins like Asf1 influencing replication-coupled chromatin assembly and cell cycle progression by modifying histones and other structural proteins.
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