FANCD2 Knockout HeLa Cell Lysate

Product Discontinued

FANCD2 Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,
• Gene: FANCD2

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for FANCD2 Knockout HeLa Cell Lysate

• DKFZp762A223
• FA4
• FACD
• FAD
• FAD2
• FA-D2
• FADFAD2
• FANCD
• FANCD2
• Fanconi anemia complementation group D2
• Fanconi anemia group D2 protein
• Fanconi anemia, complementation group D2
• FLJ23826
• FPN1
• HFE4
• IREG1
• Protein FACD2
• SLC11A3

Background

Fanconi Anemia (FA) is an autosomal-recessive cancer-prone disorder characterized by congenital defects, progressive bone marrow failure, increased chromosomal breakage, defective DNA repair and cellular hypersensitivity to mitomycin C. Fanconi Anemia Complementation Group D (FANCD) is comprised of two separate proteins, FANCD1/BRCA2 and FANCD2. LYRIC/MTDH, an RNA binding protein, regulates expression of FANCD2 and FANCD1 (1). FANCD2 is mapped to chromosome 3p25.3, has a theoretical molecular weight of 166 kDa and is localized in the nucleus. FANCD2 is involved in the regulation of DNA-binding transcription factor activity and DNA stability through accurate and efficient pairing of homologs during meiosis to promote repair of double-strand DNA breaks (2). Ubiquitination is required for FANCD2 to bind to chromatin. In response to DNA damage, FANCD2 is monoubiquinated to result in colocalization with other proteins (BRCA1 and BRCA2) involved in homology-directed DNA repair in the nucleus and is deubiquitinated upon DNA repair completion.

References

1. Bi, J., Areecheewakul, S., Li, Y., Yang, S., Zhang, Y., Ebeid, K., . . . Meng, X. (2019). MTDH/AEG-1 downregulation using pristimerin-loaded nanoparticles inhibits Fanconi anemia proteins and increases sensitivity to platinum-based chemotherapy. Gynecol Oncol, 155(2), 349-358. doi:10.1016/j.ygyno.2019.08.014

2. Balcerek, J., Jiang, J., Li, Y., Jiang, Q., Holdreith, N., Singh, B., . . . Tong, W. (2018). Lnk/Sh2b3 deficiency restores hematopoietic stem cell function and genome integrity in Fancd2 deficient Fanconi anemia. Nat Commun, 9(1), 3915. doi:10.1038/s41467-018-06380-1

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: FANCD2