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AB257434

Human FANCI knockout HEK-293T cell lysate

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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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Western blot - Human FANCI knockout HEK-293T cell lysate (AB257434)
  • WB

Lab

Western blot - Human FANCI knockout HEK-293T cell lysate (AB257434)

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.

All lanes:

Western blot - Anti-FANCI antibody (<a href='/en-us/products/primary-antibodies/fanci-antibody-ab103077'>ab103077</a>) 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 (<a href='/en-us/products/cell-lines/human-fanci-knockout-hek-293t-cell-line-ab266278'>ab266278</a>)

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

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 149 kDa

Observed band size: 151 kDa

false

Sanger Sequencing - Human FANCI knockout HEK-293T cell lysate (AB257434)
  • Sanger seq

Unknown

Sanger Sequencing - Human FANCI knockout HEK-293T cell lysate (AB257434)

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

Sanger Sequencing - Human FANCI knockout HEK-293T cell lysate (AB257434)
  • Sanger seq

Unknown

Sanger Sequencing - Human FANCI knockout HEK-293T cell lysate (AB257434)

Allele-1 : 2 bp deletion in exon 4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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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Complementary Products

Primary Antibodies

AB245219

Anti-FANCI antibody [EPR23903-124]

BOND RX™ Validated|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCI antibody [EPR23903-124] (AB245219)

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  • 0
Cell Lines & Lysates

AB273707

Human FANCI knockout HEK-293 cell line

Next Generation Sequencing - Human FANCI knockout HEK-293 cell line (AB273707)

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  • 0
Proteins & Peptides

AB132153

Recombinant Human FANCI protein

SDS-PAGE - Recombinant Human FANCI protein (AB132153)

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  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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.

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What's included?

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Properties and storage information

Gene name
FANCI
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.
Biological function summary

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.

Pathways

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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Quality control

STR analysis

TH01, D16S539, TPOX, CSF1PO, D13S317, D7S820, D5S818

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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

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