FGFR1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
Images
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout.
Recommended products
FGFR1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout.
- Disease
- Glioblastoma
- Concentration
Properties
- Gene name
- FGFR1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Viability
- ~ 80%
- Adherent/suspension
- Adherent
- Gender
- Male
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
- EMEM + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Recommended control: Human wild-type U-87 MG cell line (ab278079). 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.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
FGFR1 also known as fibroblast growth factor receptor 1 is a protein with a molecular weight of approximately 92 kDa. It is a receptor tyrosine kinase that binds to fibroblast growth factors (FGFs) triggering a cascade of downstream signaling pathways. FGFR1 is widely expressed in various tissues including the brain skeletal muscle and the cardiovascular system. The FGFR1 protein plays an essential role in cellular processes such as proliferation differentiation and survival.
- Biological function summary
FGFR1 is significant in embryonic development and tissue repair. It does not function alone; rather it forms a complex with fibroblast growth factors and heparan sulfate proteoglycans facilitating receptor dimerization and autophosphorylation. FGFR1 is involved in bone growth angiogenesis and wound healing processes. The interaction of FGFR1 with these complex factors ensures the precise regulation of these critical physiological events in the body.
- Pathways
FGFR1 participates actively in the MAPK/ERK and PI3K/AKT signaling pathways. These pathways are vital for transmitting signals from the cell surface to the DNA in the cell nucleus regulating gene expression and affecting cell cycle progression. FGFR1 works alongside similar proteins like FGFR2 influencing cell fate decisions. By modulating these pathways FGFR1 ensures a proper response to environmental signals which maintains homeostasis and adapts to physiological needs.
- Associated diseases and disorders
FGFR1 is implicated in conditions such as cancer and skeletal dysplasias. Abnormal FGFR1 signaling due to mutations or overexpression can lead to tumorigenesis contributing to the development of cancers like breast and lung cancer. The protein FGFR2 is often evaluated in parallel for similar oncogenic activities. Moreover FGFR1-related dysregulation in bone development can result in skeletal disorders highlighting its importance in both normal physiology and pathological conditions. Anti-FGFR therapies and FGFR1 ELISA tools are being explored to better understand and tackle these diseases.
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2 product images
Western blot - Human FGFR1 knockout U-87 MG cell line (ab306662)
Western blot: Anti-FGFR1 antibody [EPR806Y] (Anti-FGFR1 antibody [EPR806Y] ab76464) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-FGFR1 antibody [EPR806Y] ab76464 was shown to bind specifically to FGFR1. A band was observed at 92 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in FGFR1 knockout cell line. To generate this image, wild-type and FGFR1 knockout U-87 MG 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-FGFR1 antibody [EPR806Y] (Anti-FGFR1 antibody [EPR806Y] ab76464) at 1/500 dilution
Lane 1: Wild-type U-87 MG cell lysate at 20 µg
Lane 2: FGFR1 knockout U-87 MG cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 92 kDa
Next Generation Sequencing - Human FGFR1 knockout U-87 MG cell line (ab306662)
122 bp deletion after Gly269 of the WT protein
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