Human FYN knockout HEK-293T cell line
- Advanced Validation
- What is this?
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FYN KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion 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
C syn protooncogene, FYN oncogene related to SRC FGR YES, FYN_HUMAN, OKT3 induced calcium influx regulator, Protein tyrosine kinase fyn, Proto oncogene tyrosine protein kinase fyn, Proto-oncogene Syn, Proto-oncogene c-Fyn, SYN, Src yes related novel gene, Src-like kinase, Src/yes related novel, Tyrosine kinase p59fyn T, Tyrosine kinase p59fyn(T), Tyrosine-protein kinase Fyn, p59-Fyn
- WB
Lab
Western blot - Human FYN knockout HEK-293T cell line (AB266133)
Lanes 1-3 : Merged signal (red and green). Green - ab184276 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab184276 Anti-Fyn antibody [EPR19636] was shown to specifically react with Fyn in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266133 (knockout cell lysate ab257071) was used. Wild-type and Fyn knockout samples were subjected to SDS-PAGE. ab184276 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Fyn antibody [EPR19636] (<a href='/en-us/products/primary-antibodies/fyn-antibody-epr19636-ab184276'>ab184276</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
FYN knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human FYN knockout HEK-293T cell line (ab266133)
Lane 3:
Human brain tissue lysate at 20 µg
Predicted band size: 60 kDa
Observed band size: 159 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human FYN knockout HEK-293T cell line (AB266133)
Homozygous : 1 bp insertion in exon 4
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
Fyn kinase influences cell growth differentiation and survival. It is a part of larger signaling complexes including those accountable for the development and function of the central nervous system. The protein's activity is important for neural plasticity and supports synaptic communication particularly within the brain. Fyn can interact with other proteins in these complexes to help direct cellular responses to external signals.
Pathways
Fyn kinase is central to several intracellular signaling pathways such as the T cell receptor (TCR) signaling pathway and the integrin signaling pathway. The TCR signaling pathway involves the regulation of immune responses while the integrin signaling pathway contributes to cell adhesion and migration. Other proteins like Lck and Yes interact with Fyn within these pathways adding another layer of regulation and response in various cell types.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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