FANCI KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.
FANCI gene, FANCI_HUMAN, FLJ10719, FLJ14658, Fanconi anemia group I protein, Fanconi anemia, complementation group I, KIAA1794, Protein FACI, Protein FANCI
FANCI KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
FANCI also known as Fanconi anemia group I protein is an important player in DNA repair processes. This protein has a molecular mass of approximately 150 kDa. It is ubiquitously expressed with notable levels found in tissues that undergo high rates of cell division. FANCI participates mechanically within the DNA repair mechanism by interacting directly with DNA alongside its partner protein FANCD2 to form a tight complex that recognizes and binds to sites of DNA damage.
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.
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.
FANCI mutations are directly linked to Fanconi anemia a genetic disorder characterized by bone marrow failure and increased cancer risk. Mutations can impair the protein’s function causing defective DNA repair capability. Additionally FANCI's involvement in DNA repair associates it with breast cancer where malfunction or reduced expression of FANCI is observed. The protein interacts with tumor suppressors like BRCA1 and BRCA2 linking it to pathways involved in hereditary breast cancer predisposition.
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Anti-FANCI antibody ab103077 was shown to specifically react with FANCI in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human FANCI knockout HEK-293T cell line ab266278 (knockout cell lysate ab257434) was used. Wild-type and FANCI knockout samples were subjected to SDS-PAGE. Anti-FANCI antibody ab103077 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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 (Anti-FANCI antibody ab103077) 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 (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
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 149 kDa
Observed band size: 151 kDa
Allele-1: 2 bp deletion in exon 4
Allele-2: Insertion of the selection cassette in exon 4
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