FZR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 9.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 9
Alternative names
CDC20 like 1b, CDC20-like protein 1, CDC20C, CDH1, Cdh1/Hct1 homolog, FYR, FZR 2, FZR_HUMAN, Fizzy related protein, Fizzy related protein 1, Fizzy-related protein homolog, Fizzy/cell division cycle 20 related 1, Fzr, Fzr1 protein, HCDH 1, KIAA1242
Recommended products
FZR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 9.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 9
- Concentration
Properties
- Gene name
- FZR1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
FZR1 also known as fizzy-related protein homolog plays a critical mechanical role in cell cycle regulation as a co-activator of the anaphase-promoting complex/cyclosome (APC/C). The FZR1 protein has a mass of about 83 kDa. It is involved in the transition from metaphase to anaphase during cell division. This protein expresses in various tissues but shows higher levels in proliferating cells due to its role in cell cycle control.
- Biological function summary
FZR1 helps regulate the degradation of cell cycle proteins by tagging them for destruction in a process involving the ubiquitin-proteasome system. FZR1 forms a complex with APC/C pushing the progression of the cell cycle and ensuring proper chromosome segregation. This activity prevents genomic instability by maintaining the correct order of cell cycle phases.
- Pathways
Researchers place FZR1 in the cell cycle pathway particularly in regulating the mitotic exit and G1/S transition. One significant pathway involves its partnership with the cyclin B1 protein where FZR1 aids in its ubiquitination and subsequent degradation. It is also related to pathways involving the retinoblastoma protein (Rb) which adds a layer of regulation over the cycle and ensures DNA replication precision.
- Associated diseases and disorders
FZR1 associates with cancer due to its involvement in controlling cell division and preventing aneuploidy. Disruption in FZR1 function can lead to misregulated cell growth which is often observed in tumors. Furthermore some studies connect FZR1 to neurodegenerative disorders notably through its interactions with proteins in the nervous system although these connections are still under investigation.
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3 product images
Western blot - Human FZR1 knockout HEK-293T cell line (ab267263)
Lanes 1-4: Merged signal (red and green). Green - Anti-FZR1 antibody [AR38.2] ab77885 observed at 55 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 observed at 36 kDa.Anti-FZR1 antibody [AR38.2] ab77885 Anti-FZR1 antibody [AR38.2] was shown to specifically react with FZR1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab267263 (knockout cell lysate Human FZR1 knockout HEK-293T cell lysate ab263208) was used. Wild-type and FZR1 knockout samples were subjected to SDS-PAGE. Anti-FZR1 antibody [AR38.2] ab77885 and Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-FZR1 antibody [AR38.2] (Anti-FZR1 antibody [AR38.2] ab77885) at 1/500 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: FZR1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: Western blot - Human FZR1 knockout HEK-293T cell line (ab267263)
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
Cell Culture - Human FZR1 knockout HEK-293T cell line (ab267263)
Representative images FZR1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Sanger Sequencing - Human FZR1 knockout HEK-293T cell line (ab267263)
Homozygous: 1 bp insertion in exon9
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