Product Pathways - Chromatin Regulation / Epigenetics
5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb #74178
|74178S||100 µl ( 10 Dot Blots )||￥3,100.00 现货查询||购买询价|
|74178||carrier free & custom formulation / quantity||email request|
|IF-IC||1:200||All Species Expected,||Endogenous||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry), Dot Blot=DNA Dot Blot,
Specificity / Sensitivity
5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb recognizes endogenous levels of 5-formylcytosine. This antibody has been validated for specificity using ELISA and dot blot and shows high specificity for 5-formylcytosine.
Source / Purification
Monoclonal antibody is produced by immunizing animals with 5-formyl-2'-deoxycytosine.
5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb specificity was determined by ELISA. The antibody was titrated against a single-stranded DNA oligo containing either unmodified cytosine or differentially modified cytosine (5-mC, 5-hmC, 5-caC, 5-fC). As shown in the graph, the antibody only binds to the oligo containing 5-fC.
The specificity of 5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb was determined by dot blot. The same sequence of a 387-base pair DNA fragment was generated by PCR using exclusively unmodified cytosine, 5-methylcytosine (5-mC), 5-hydroxymethylcytosine (5-hmC), 5-carboxylcytosine (5-caC), or 5-formylcytosine (5-fC). The respective DNA fragments were blotted onto a nylon membrane, UV cross-linked, and probed with 5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb. The upper panel shows the antibody only binding to the DNA fragment containing 5-fC, while the lower panel shows the membrane stained with methylene blue.
Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing DDK-tagged TET1 catalytic domain (TET1-CD) using 5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb (green) and DYKDDDDK Tag (9A3) Mouse mAb #8146 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). As expected, 293T cells expressing TET1-CD (red) exhibit increased levels of 5-formylcytosine (green).
Methylation of DNA at cytosine residues is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and mammalian development (1,2). 5-methylcytosine is a repressive epigenetic mark established de novo by two enzymes, DNMT3a and DNMT3b, and is maintained by DNMT1 (3, 4). 5-methylcytosine was originally thought to be passively depleted during DNA replication. However, subsequent studies have shown that Ten-Eleven Translocation (TET) proteins TET1, TET2, and TET3 can catalyze the oxidation of methylated cytosine to 5-hydroxymethylcytosine (5-hmC) (5). Additionally, TET proteins can further oxidize 5-hmC to form 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC), both of which are excised by thymine-DNA glycosylase (TDG), effectively linking cytosine oxidation to the base excision repair pathway and supporting active cytosine demethylation (6,7).
TET protein-mediated cytosine hydroxymethylation was initially demonstrated in mouse brain and embryonic stem cells (5, 8). Since then this modification has been discovered in many tissues, with the highest levels found in the brain (9). While 5-fC and 5-caC appear to be short-lived intermediate species, there is mounting evidence showing that 5-hmC is a distinct epigenetic mark with various unique functions (10,11). The modified base itself is stable in vivo and interacts with various readers including MeCP2 (11,12). The global level of 5-hmC increases during brain development, and 5-hmC is enriched at promoter regions and poised enhancers. Furthermore, there is an inverse correlation between levels of 5-hmC and histone H3K9 and H3K27 trimethylation, suggesting a role for 5-hmC in gene activation (12). Lower amounts of 5-hmC have been reported in various cancers including myeloid leukemia and melanoma (13,14).
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- Globisch, D. et al. (2010) PLoS One 5, e15367.
- Gao, Y. et al. (2013) Cell Stem Cell 12, 453-69.
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Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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For Research Use Only. Not For Use In Diagnostic Procedures.
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