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Human FANCI knockout HEK293T cell pellet (ab278872)

Overview

  • Product name

    Human FANCI knockout HEK293T cell pellet
    See all FANCI kits

  • Product overview

    Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.

    Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation.

  • Parental Cell Line

    HEK293T

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.

  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Notes

    Pellet size: 5 million cells/vial.

    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.

  • Tested applications

    Suitable for: WBmore details

Properties

Target

  • Function

    Required for maintenance of chromosomal stability. Involved in the repair of DNA double-strand breaks by homologous recombination and in the repair of DNA cross-links. Participates in S phase and G2 phase checkpoint activation upon DNA damage. Promotes FANCD2 ubiquitination and recruitment to DNA repair sites.

  • Involvement in disease

    Defects in FANCI are a cause of Fanconi anemia complementation group I (FANCI) [MIM:609053]. It is a disorder affecting all bone marrow elements and resulting in anemia, leukopenia and thrombopenia. It is associated with cardiac, renal and limb malformations, dermal pigmentary changes, and a predisposition to the development of malignancies. At the cellular level it is associated with hypersensitivity to DNA-damaging agents, chromosomal instability (increased chromosome breakage) and defective DNA repair.

  • Domain

    The C-terminal 30 residues are probably required for function in DNA repair.

  • Post-translational
    modifications

    Monoubiquitinated on Lys-523 during S phase and upon genotoxic stress. Deubiquitinated by USP1 as cells enter G2/M, or once DNA repair is completed. Monoubiquitination requires the FANCA-FANCB-FANCC-FANCE-FANCF-FANCG-FANCM complex. Ubiquitination is required for binding to chromatin, DNA repair, and normal cell cycle progression.
    Phosphorylated in response to DNA damage by ATM and/or ATR.

  • Cellular localization

    Nucleus. Concentrates in nuclear foci upon genotoxic stress.
  • Information by UniProt

  • Alternative names

    • FANCI
    • FANCI gene
    • FANCI_HUMAN
    • Fanconi anemia group I protein
    • Fanconi anemia, complementation group I
    • FLJ10719
    • FLJ14658
    • KIAA1794
    • Protein FACI
    • Protein FANCI
    see all

Properties

Images

  • Western blot - Human FANCI knockout HEK293T cell pellet (ab278872)
    Western blot - Human FANCI knockout HEK293T cell pellet (ab278872)

    Lane 1:Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
    Lane 2:FANCI knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
    Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
    Lane 4:Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)

    ab103077 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 4oC 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.

  • Sanger Sequencing - Human FANCI knockout HEK293T cell pellet (ab278872)
    Sanger Sequencing - Human FANCI knockout HEK293T cell pellet (ab278872)

    Allele-1: 2 bp deletion in exon 4

     

  • Sanger Sequencing - Human FANCI knockout HEK293T cell pellet (ab278872)
    Sanger Sequencing - Human FANCI knockout HEK293T cell pellet (ab278872)

    Allele-2: Insertion of the selection cassette in exon 4

     

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