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AB269630

Human FYN knockout HEK-293 cell line

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FYN KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%. 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

3 Images
Western blot - Human FYN knockout HEK-293 cell line (AB269630)
  • WB

Lab

Western blot - Human FYN knockout HEK-293 cell line (AB269630)

False colour image of Western blot : Anti-Fyn antibody [EPR19636] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab184276 was shown to bind specifically to Fyn. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in FYN knockout cell line ab269630 (knockout cell lysate ab272440). To generate this image, wild-type and FYN knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. 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-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-293 cell lysate at 20 µg

Lane 2:

FYN knockout HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human FYN knockout HEK-293 cell line (ab269630)

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Western blot - Human FYN knockout HEK-293 cell line (AB269630)
  • WB

Lab

Western blot - Human FYN knockout HEK-293 cell line (AB269630)

False colour image of Western blot : Anti-Fyn antibody [EPR5500] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab125016 was shown to bind specifically to Fyn. A band was observed at 60 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in FYN knockout cell line ab269630 (knockout cell lysate ab272440). To generate this image, wild-type and FYN knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. 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-Fyn antibody [EPR5500] (<a href='/en-us/products/primary-antibodies/fyn-antibody-epr5500-ab125016'>ab125016</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

FYN knockout HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human FYN knockout HEK-293 cell line (ab269630)

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Next Generation Sequencing - Human FYN knockout HEK-293 cell line (AB269630)
  • NGS

Supplier Data

Next Generation Sequencing - Human FYN knockout HEK-293 cell line (AB269630)

Knockout achieved by CRISPR/Cas9; X = 14 bp deletion; Frameshift : 100%

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 14 bp deletion Frameshift: 100%

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
FYN
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Fyn is a Src family tyrosine kinase with a molecular weight of approximately 59 kDa. It is often referred to as Fyn kinase and plays a significant role in signal transduction processes. This protein is observed in various tissues with high expression in the brain T cells and platelets. Fyn kinase is known for its function in cell communication and regulates different cellular processes in these tissues.
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.

Fyn kinase has associations with Alzheimer's disease and certain types of cancer. In Alzheimer's dysregulation of Fyn activity contributes to synaptic dysfunction and neuronal damage. In cancer aberrant Fyn signaling can affect cellular proliferation and metastasis. Proteins such as Tau in Alzheimer's disease and Src in cancer relate to Fyn through their respective pathological processes.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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