Human FUS (TLS) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human FUS (TLS) knockout HEK-293T cell line (AB266587)
Lane 1 : Wild-type HEK-293T cell lysate (20µg)
Lane 2 : FUS knockout HEK-293T cell lysate (20µg)
Lanes 1- 2 : Merged signal (red and green). Green - ab124923 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124923 Anti-TLS/FUS antibody [EPR5812] was shown to specifically react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type and TLS/FUS 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. ab124923 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TLS/FUS antibody [EPR5812] (<a href='/en-us/products/primary-antibodies/tls-fus-antibody-epr5812-ab124923'>ab124923</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human FUS (TLS) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-fus-tls-knockout-hek-293t-cell-lysate-ab257100'>ab257100</a>) at 20 µg
Secondary
Lanes 1 - 2:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 75 kDa,37 kDa
false
- WB
Lab
Western blot - Human FUS (TLS) knockout HEK-293T cell line (AB266587)
Lanes 1- 2 : Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab154141 was shown to react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type HEK-293T and FUS knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154141 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TLS/FUS antibody [CL0190] (<a href='/en-us/products/primary-antibodies/tls-fus-antibody-cl0190-ab154141'>ab154141</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
FUS knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human FUS (TLS) knockout HEK-293T cell line (ab266587)
Predicted band size: 53 kDa
Observed band size: 80 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human FUS (TLS) knockout HEK-293T cell line (AB266587)
Homozygous : 1 bp deletion in exon2
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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 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?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Quality control - STR analysis
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
The FUS protein aids in the maintenance and stabilization of mRNA molecules and participates in the formation of stress granules distinct cytoplasmic aggregates of proteins and RNAs. It forms part of a large protein complex alongside other RNA-binding proteins. FUS stabilizes pre-mRNA structures which affects protein expression patterns essential for cell survival and differentiation. Its ability to bind to RNA and DNA indicates its fundamental role in genomic stability.
Pathways
The functions of FUS help regulate RNA metabolism-related pathways and cellular stress response pathways. FUS interacts with proteins like TAF15 and EWSR1 within these pathways showing a complex network of interactions that contribute to RNA maturation and stress granule dynamic formation. The activity of FUS in these pathways ultimately affects cellular homeostasis gene expression modulation and response to cellular stress.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
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