Human FANCI knockout HEK-293T cell line
- Advanced Validation
- What is this?
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FANCI KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.
View Alternative Names
FANCI gene, FANCI_HUMAN, FLJ10719, FLJ14658, Fanconi anemia group I protein, Fanconi anemia, complementation group I, KIAA1794, Protein FACI, Protein FANCI
- WB
Lab
Western blot - Human FANCI knockout HEK-293T cell line (AB266278)
Lanes 1-4 : Merged signal (red and green). Green - ab103077 observed at 151 kDa. Red - loading control ab8245 observed at 36 kDa.
ab103077 Anti-FANCI antibody was shown to specifically react with FANCI in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266278 (knockout cell lysate ab257434) was used. Wild-type and FANCI knockout samples were subjected to SDS-PAGE. ab103077 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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® 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-FANCI antibody (<a href='/en-us/products/primary-antibodies/fanci-antibody-ab103077'>ab103077</a>) 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:
FANCI 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 FANCI knockout HEK-293T cell line (ab266278)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Raji (Human Burkitts lymphoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
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 1/10000 dilution
Predicted band size: 149 kDa
Observed band size: 151 kDa
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- Cell Culture
Lab
Cell Culture - Human FANCI knockout HEK-293T cell line (AB266278)
Representative images FANCI 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 seq
Unknown
Sanger Sequencing - Human FANCI knockout HEK-293T cell line (AB266278)
Allele-2 : Insertion of the selection cassette in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human FANCI knockout HEK-293T cell line (AB266278)
Allele-1 : 2 bp deletion in exon 4
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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.
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
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
The FANCI protein acts as part of the multiprotein Fanconi anemia (FA) core complex. This complex Functions in the repair of DNA interstrand crosslinks which are severe forms of DNA damage. Successful function of FANCI is important for maintaining genomic stability and subsequently proper cell cycle progression. It partners with proteins including FANCD2 to coordinate the homologous recombination repair process. Defective FANCI leads to disrupted DNA repair resulting in cellular damage and apoptosis.
Pathways
FANCI plays a major role in the Fanconi anemia pathway and the broader DNA damage response system. The FA pathway which involves over 20 proteins forms a network integral in repairing DNA crosslinks. FANCI tightly interacts with proteins like FANCD2 to initiate DNA repair in response to damage. The successful coordination of these proteins ensures the stability of genetic material throughout the cell cycle affecting cell fate decisions such as survival or apoptosis.
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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Cell Lines & Lysates
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Advanced Validation
- 0
- 0
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