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#97763 DNA Cytosine Modification Antibody Sampler Kit

 
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希望納入価格 (円)
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2019年1月17日15時10分 現在
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#97763T1 Kit75,000
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キット内容 容量 用途 種交差性 検出分子量 アイソタイプ
5-Methylcytosine (5-mC) (D3S2Z) Rabbit mAb #28692 20 µl IF-IC, Dot Blot, MeDIP All Rabbit IgG
5-Hydroxymethylcytosine (5-hmC) (HMC31) Mouse mAb #51660 20 µl IF-IC, Dot Blot, MeDIP All Mouse IgG1
5-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb #36836 20 µl IF-IC, Dot Blot All Rabbit IgG
5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb #74178 20 µl IF-IC, Dot Blot All Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl WB Horse

Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry), Dot Blot=DNA Dot Blot, MeDIP=Methylated DNA IP, W=Western Blotting
Reactivity Key: All=All Species Expected

貯法
-20℃
社内データ

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

IF-IC

IF-IC

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).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing DYKDDDDK-tagged TET1 catalytic domain (TET1-CD) using 5-Carboxylcytosine (5-caC) (D7S8U) 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-carboxylcytosine (green).


IF-IC

IF-IC

Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing DDK-tagged TET1 catalytic domain (TET1-CD) using 5-Hydroxymethylcytosine (5-hmC) (HMC31) Mouse mAb (green) and DYKDDDDK Tag Antibody #2368 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). As expected, 293T cells expressing TET1-CD (red) exhibit inccreased levels of 5-hydroxymethylcytosine (green).

Dot Blot-DNA

Dot Blot-DNA

Specificity of various modified cytosine antibodies were 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-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb #36836, 5-Methylcytosine (5-mC) (D3S2Z) Rabbit mAb #28692, 5-Hydroxymethylcytosine (5-hmC) (HMC31) Mouse mAb #51660, and 5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb #74178. As shown, the 5-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb shows specificity for 5-caC.

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293T cells transfected with a construct expressing DYKDDDDK-tagged TET1 catalytic domain (TET1-CD) using 5-Methylcytosine (5-mC) (D3S2Z) 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 decreased levels of 5-methylcytosine (green).


バックグラウンド

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).

使用例
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関連製品
28692   5-Methylcytosine (5-mC) (D3S2Z) Rabbit mAb
36836   5-Carboxylcytosine (5-caC) (D7S8U) Rabbit mAb
51660   5-Hydroxymethylcytosine (5-hmC) (HMC31) Mouse mAb
7003   20X LumiGLO® Reagent and 20X Peroxide
7072   Phototope-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
7076   Anti-mouse IgG, HRP-linked Antibody
74178   5-Formylcytosine (5-fC) (D5D4K) Rabbit mAb
9999   Nonfat Dry Milk

DRAQ5 is a registered trademark of Biostatus Limited.
Tween is a registered trademark of ICI Americas, Inc.
SimpleDIP is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

DNA Cytosine Modification Antibody Sampler Kit

Metabolic Reprogramming in Disease

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