Human FANCA knockout A549 cell line
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FANCA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
FA, FA 1, FA H, FAA, FACA, FANCA_HUMAN, FANCH, Fanconi anemia complementation group A, Fanconi anemia complementation group H, Fanconi anemia group A protein, Fanconi anemia type 1, MGC75158, Protein FACA
- WB
Lab
Western blot - Human FANCA knockout A549 cell line (AB287410)
Western blot : Anti-FANCA antibody [EPR16519] (ab201457) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab201457 was shown to bind specifically to FANCA. A band was observed at 163 kDa in wild-type A549 cell lysates with no signal observed at this size in FANCA knockout cell line. To generate this image, wild-type and FANCA knockout A549 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.
Lanes 1 - 4:
Western blot - Anti-FANCA/FAA antibody [EPR16519] (<a href='/en-us/products/primary-antibodies/fanca-faa-antibody-epr16519-ab201457'>ab201457</a>) at 1/1000 dilution
Lanes 1 - 4:
Western blot - Anti-FANCA/FAA antibody [EPR16519] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/fanca-faa-antibody-epr16519-bsa-and-azide-free-ab251344'>ab251344</a>)
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
FANCA knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HAP1 cell lysate at 20 µg
Lane 4:
FANCA knockout HAP1 cell lysate at 20 µg
Observed band size: 163 kDa
false
- Sanger seq
Project
Sanger Sequencing - Human FANCA knockout A549 cell line (AB287410)
59bp deletion after Val 109 of the WT protein.
- NGS
Supplier Data
Next Generation Sequencing - Human FANCA knockout A549 cell line (AB287410)
59 bp deletion after Val108 of the WT protein
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab288558). 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 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.
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
FANCA works as a part of the Fanconi anemia (FA) core complex involving around 13 proteins. This complex plays an important role in the FA DNA repair pathway facilitating the repair of interstrand DNA crosslinks. FANCA acts by orchestrating the recruitment and activation of downstream repair proteins which ensures the high-fidelity maintenance of the genetic material during cell division. Dysfunction in the FA complex where FANCA operates can lead to chromosomal instability and increased mutation rates.
Pathways
FANCA functions prominently in the Fanconi anemia pathway and is also involved in the homologous recombination repair pathway. In the FA pathway FANCA works closely with related proteins like FANCB and FANCC to initiate the DNA repair process. FANCA's interaction with these proteins allows it to execute its repair role effectively ensuring cellular genomic integrity. These pathways are critical for protecting cells from DNA damage during replication and shielding them from cancer development.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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