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FANCD2 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: 4 bp deletion in exon 13.

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Images

Western blot - Human FANCD2 knockout HeLa cell line (AB261743), expandable thumbnail
  • Western blot - Human FANCD2 knockout HeLa cell line (AB261743), expandable thumbnail
  • Sanger Sequencing - Human FANCD2 knockout HeLa cell line (AB261743), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 13

Alternative names

DKFZp762A223, FA 4, FA D2, FACD, FACD2_HUMAN, FAD, FANCD, FANCONI ANEMIA COMPLEMENTATION GROUP D, FANCONI PANCYTOPENIA TYPE 4, FLJ23826, Fanconi anemia complementation group D2, Fanconi anemia group D2 protein, OTTHUMP00000158853, OTTHUMP00000207925, Protein FACD2, Type 4 Fanconi pancytopenia

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FANCD2 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: 4 bp deletion in exon 13.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 13
Antibiotic resistance
Puromycin 1µg/mL
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
FANCD2
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

Recommended control: Human wild-type HeLa cell line (ab255448). 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

FANCD2 also known as Fanconi anemia group D2 functions mechanically in DNA repair processes. This protein plays a role in the cellular response to DNA damage specifically interstrand crosslinks. FANCD2 has a molecular weight of approximately 164 kDa. Expression of FANCD2 occurs broadly in various tissues but reaches higher levels in cells that undergo rapid division such as hematopoietic cells.

Biological function summary

FANCD2 collaborates with multiple proteins as part of the Fanconi anemia (FA) complex to ensure genomic stability. The FA complex operates by facilitating the repair of DNA interstrand crosslinks that impede replication. FANCD2 becomes activated through monoubiquitination which serves as a signal for recruitment to DNA damage sites. It essentially functions as a coordinator that brings together important components for repair.

Pathways

FANCD2 integrates into the Fanconi anemia pathway and the homologous recombination repair pathway. These pathways are critical for maintaining DNA integrity and preventing chromosomal instability. Within these pathways interactions with proteins such as BRCA1 and BRCA2 reinforce the repair processes. FANCD2's connection to these proteins exemplifies its role in complex mechanisms that preserve genomic fidelity.

Associated diseases and disorders

FANCD2 links significantly to Fanconi anemia a genetic disorder that causes bone marrow failure and increased cancer susceptibility. Damage or malfunction of FANCD2 can disrupt DNA repair leading to cellular dysfunction seen in this condition. Additionally there is a connection to breast cancer whereby FANCD2 interacts with BRCA2 indicating a shared pathway involved in tumor suppressor functions. These associations underline the importance of FANCD2 in disease pathology and its potential as a therapeutic target.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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