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验证数据展示

  • ELISA experiment of 5-Hydroxymethylcytidine using 68579-1-Ig

    ELISA experiment of 5-Hydroxymethylcytidine using 68579-1-Ig

    Indirect ELISA and competitive ELISA results show that this antibody is specific to 5-Hydroxymethylcytidine (hm5C). Indirect ELISA was performed by coating BSA conjugated 5-Hydroxymethylcytidine (hm5C) at 10ng/well followed by blocking with 1% BSA. Serial diluted primary antibody was added to the plates and incubated at 37℃. HRP-goat anti-mouse was used for detection. Competitive ELISA was performed similarly except that different concentration of 5-Hydroxymethylcytidine (hm5C) or its structure analogue compounds are mixed in primary antibody (fixed at 10 ng/mL).

  • Dot Blot experiment of RNA using 68579-1-Ig

    Dot Blot experiment of RNA using 68579-1-Ig

    Total RNA was isolated from HeLa cell line and was dotted to NC membrane at different amount as indicated above the dots. The membrane was blocked with BSA and blotted with 5-Hydroxymethylcytidine (hm5C) antibody 68579-1-Ig at 1:5000 followed by incubation of HRP-goat anti-mouse secondary antibody. Signal was developed by ECL substrate. A parallel dot blot was performed using mouse IgG1 isotype control antibody 66360-1-Ig at the same dose.

5-Hydroxymethylcytidine Monoclonal antibody

5-Hydroxymethylcytidine Monoclonal Antibody for Dot Blot, ELISA
Cat No. 68579-1-Ig

产品说明书

CloneNo. 1E6C6

宿主/亚型

Mouse / IgG1

种属反应性

chemical compound

应用

Dot Blot, ELISA

别名

hm5C

缓冲液配方:  PBS and Azide
PBS and Azide
PBS Only
偶联物:  Unconjugated
Unconjugated
规格: 

-/ -

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经过测试的应用

Positive ELISA detected in5-Hydroxymethylcytidine
Positive Dot Blot detected inRNA

推荐稀释比

应用推荐稀释比
Enzyme-linked Immunosorbent Assay (ELISA)ELISA : 1:2000-1:20000
DOT BLOTDOT BLOT : 1:1000-1:4000
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

产品信息

68579-1-Ig targets 5-Hydroxymethylcytidine in Dot Blot, ELISA applications and shows reactivity with chemical compound samples.

经测试应用 Dot Blot, ELISA Application Description
经测试反应性 chemical compound
免疫原 PTG 种属同源性预测
宿主/亚型 Mouse / IgG1
抗体类别 Monoclonal
产品类型 Antibody
全称 5-Hydroxymethylcytidine
别名 hm5C
基因名称
Gene ID (NCBI)
RRIDAB_3085279
偶联类型 Unconjugated
形式 Liquid
纯化方式Protein G purification
储存缓冲液 PBS with 0.02% sodium azide and 50% glycerol , pH 7.3
储存条件Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

背景介绍

Oxidation of 5-methylcytosine in DNA by ten-eleven translocation (Tet) family of enzymes has been demonstrated to play a significant role in epigenetic regulation in mammals. Recent reaearch shows that Tet enzymes also possess the activity of catalyzing the formation of 5-hydroxymethylcytidine (5-hmrC) in RNA. It is known that RNA carries more than 100 distinct types of modifications, and these modifications modulate the structure and functions of RNA. Ribonucleoside 5-methylcytidine (m5C) is subject to oxidative processing in mammals, forming 5-hydroxymethylcytidine (hm5C) and 5-formylcytidine (f5C). Researchers have identified hm5C in total RNA from all three domains of life and in polyA-enriched RNA fractions from mammalian cells. This suggests m5C oxidation is a conserved process that could have critical regulatory functions inside cells (PMID: 25676849).

Protocol for Dot Blot:
https://www.ptglab.com/protocol/68579-1-IgDotBlot.pdf