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5-Hydroxymethylcytidine Monoclonal antibody

5-Hydroxymethylcytidine Monoclonal Antibody for Dot Blot, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human

Applications

Dot Blot, ELISA

Conjugate

Unconjugated

CloneNo.

1E6C6

Cat No : 68579-1-Ig

Print datasheet

Synonyms

5-Hydroxymethylcytidine, hm5C

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.

经过测试的应用

Positive ELISA detected in5-Hydroxymethylcytidine
Positive Dot Blot detected inRNA

推荐稀释比

ApplicationDilution
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 ELISA, Dot Blot applications and shows reactivity with Human samples.

Tested Applications Dot Blot, ELISA
Tested Reactivity Human
Immunogen PTG 种属同源性预测
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Full Name 5-Hydroxymethylcytidine
Synonyms 5-Hydroxymethylcytidine, hm5C
Gene Symbol
Gene ID (NCBI)
Conjugate Unconjugated
Form Liquid
Purification MethodProtein G purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore 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).