Human TYK2 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human TYK2 knockout HEK-293T cell line (AB266730)
False colour image of Western blot : Anti-TYK2 antibody staining at 0.04 µg/ml shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab223733 was shown to bind specifically to TYK2. A band was observed at 133 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in TYK2 knockout cell line ab266730 (knockout cell lysate ab258743). To generate this image wild-type and TYK2 knockout HEK-293T cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-TYK2 antibody (<a href='/en-us/products/primary-antibodies/tyk2-antibody-ab223733'>ab223733</a>) at 0.04 µg/mL
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
TYK2 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human TYK2 knockout HEK-293T cell line (ab266730)
Lane 3:
MCF7 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Predicted band size: 134 kDa
Observed band size: 133 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TYK2 knockout HEK-293T cell line (AB266730)
Homozygous : 1 bp insertion in exon 4
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Reactivity data
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
TYK2 mediates intracellular signaling that affects both innate and adaptive immune responses. It associates with cytokine receptors forming functional complexes that are critical for initiating JAK-STAT signaling. This type of signaling is responsible for regulating gene expression after cytokine binding. TYK2 is known for interacting with Type I interferons and interleukin-10 family cytokines indicating its importance in immune regulation.
Pathways
TYK2 acts within the JAK-STAT pathway a fundamental signaling cascade triggered by various cytokines and growth factors. The JAK-STAT pathway includes key partners such as STAT1 STAT2 and STAT3 proteins which are phosphorylated by TYK2 to propagate signal transduction. Additionally TYK2 is involved in the 'intercept blocking buffer' mechanism that adjusts signal strength thereby ensuring appropriate cellular responses to external stimuli.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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