FAN1 Antibody
Description
Applications in Research
FAN1 antibodies enable precise detection of FAN1 in diverse experimental contexts:
Western Blotting
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Validation: ab68572 detects FAN1 in human spleen lysates and transfected 293T cells, with no signal in non-transfected controls .
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Sensitivity: Detects FAN1 at dilutions as low as 1/500 in WB .
Immunoprecipitation (IP)
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Efficiency: ab95171 successfully immunoprecipitates FAN1 from HeLa lysates, confirmed via chemiluminescent detection .
Immunohistochemistry (IHC-P)
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Tissue Analysis: ab95171 labels FAN1 in formalin-fixed paraffin-embedded ovarian carcinoma tissues, enabling spatial localization studies .
Role in FAN1-Related Research
FAN1 antibodies have facilitated studies on FAN1’s function in DNA repair and repeat expansion diseases.
DNA Repair Pathways
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ICL Repair: FAN1 interacts with ubiquitinated FANCD2/FANCI complexes at DNA damage sites, as shown in mitomycin C (MMC)-treated cells . Antibodies have been used to confirm FAN1’s recruitment to damage foci .
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Repeat Expansion Modulation: FAN1 suppresses CAG/CTG repeat expansions in Huntington’s disease (HD) and fragile X syndromes. Antibodies help quantify FAN1 levels in knockout models, linking nuclease activity to expansion suppression .
Cancer and Disease Associations
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Colorectal Cancer: Germline FAN1 mutations impair DNA repair, increasing cancer risk. Antibodies aid in diagnosing FAN1 deficiency in patient samples .
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Neurodegeneration: FAN1 stabilizes expanded CAG repeats in HD. Studies using antibodies demonstrate that FAN1 knockdown accelerates expansion rates in iPSCs .
Table 1: FAN1 Antibody Performance in Key Studies
Critical Insights
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Nuclease Dependency: Mutations in FAN1’s nuclease domain (e.g., D963A) abolish its protective effects against repeat expansions, as shown via WB and expansion assays .
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FANCD2 Independence: FAN1’s repeat-suppression activity does not require FANCD2, as confirmed in Fancd2 knockout models .
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Additive Effects with EXO1: FAN1 and EXO1 nucleases synergistically suppress expansions, suggesting complementary repair mechanisms .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
What is the role of FAN1 in DNA repair, and how does it relate to the use of FAN1 antibodies in research?
FAN1 is a nuclease involved in the repair of DNA interstrand crosslinks (ICLs) and plays a role in maintaining genomic stability. It is recruited to sites of DNA damage by interacting with the monoubiquitinated FANCI-FANCD2 complex. FAN1 antibodies are used to study its expression and localization in various cell types, which is crucial for understanding its function in DNA repair processes. For example, Western blot and immunohistochemistry can be employed to assess FAN1 protein levels in different tissues or cell lines .
How does FAN1 interact with other DNA repair proteins, and what implications does this have for experimental design?
FAN1 interacts with mismatch repair proteins like MLH1, MLH3, and PMS2. These interactions suggest that FAN1 may also play a role in the repair of other types of DNA damage beyond ICLs. When designing experiments, researchers should consider co-immunoprecipitation assays to study these interactions and their impact on DNA repair pathways .
What are the implications of FAN1 mutations for research on repeat expansion diseases?
Mutations in FAN1 have been linked to karyomegalic interstitial nephritis and may influence the progression of repeat expansion diseases like Huntington's disease. Research using FAN1 antibodies can help elucidate how FAN1 variants affect disease onset and progression by analyzing protein expression in patient-derived cells .
How does FAN1's nuclease activity impact its protective role against repeat expansions, and what experimental approaches can be used to study this?
FAN1's nuclease activity is crucial for protecting against repeat expansions in some models, while in others, its protective effects may be nuclease-independent. Researchers can use site-directed mutagenesis to create nuclease-deficient FAN1 variants and assess their impact on repeat stability in cell models .
What are the best practices for validating FAN1 antibody specificity and sensitivity in research applications?
Validating antibody specificity involves using multiple antibodies from different sources and confirming results across different applications (e.g., Western blot, immunohistochemistry). Sensitivity can be assessed by titrating the antibody concentration and comparing signals across different cell types or tissues .
How can FAN1 antibodies be used to study the relationship between FAN1 expression and disease progression in Huntington's disease models?
FAN1 antibodies can be used to monitor FAN1 protein levels in Huntington's disease models, such as patient-derived iPSCs or mouse models. This can help determine if changes in FAN1 expression correlate with disease progression or CAG repeat expansion rates .
What are the potential challenges and limitations of using FAN1 antibodies in research, particularly in complex biological systems?
Challenges include ensuring antibody specificity and sensitivity across different tissues and cell types. Additionally, interpreting results in complex systems requires careful consideration of potential interactions with other DNA repair proteins and pathways .
How can researchers integrate FAN1 antibody data with other experimental approaches to gain a comprehensive understanding of FAN1's role in DNA repair and disease?
Integrating FAN1 antibody data with other techniques like CRISPR gene editing, RNA interference, and biochemical assays can provide a comprehensive view of FAN1's function. This multi-faceted approach helps elucidate its role in DNA repair and its impact on disease progression .
What are some emerging trends or future directions in the use of FAN1 antibodies for studying DNA repair mechanisms?
Emerging trends include using FAN1 antibodies in combination with advanced imaging techniques to study DNA repair dynamics in real-time and exploring FAN1's role in other diseases beyond repeat expansion disorders .
How can researchers address potential data contradictions when using FAN1 antibodies across different experimental systems or studies?
Addressing data contradictions involves carefully comparing experimental conditions, antibody specificity, and cell or tissue types used across studies. Additionally, validating findings through independent replication and using multiple experimental approaches can help resolve discrepancies .
