Anticorps Monoclonal anti-5-Hydroxymethylcytidine

5-Hydroxymethylcytidine Monoclonal Antibody for Dot Blot, ELISA

Hôte / Isotype

Mouse / IgG1

Réactivité testée

chemical compound

Applications

Dot Blot, ELISA

Conjugaison

Non conjugué

CloneNo.

1E6C6

N° de cat : 68579-1-Ig

Synonymes

hm5C

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Galerie de données de validation

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  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).

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  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.

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Applications testées

• Résultats positifs en ELISA: 5-Hydroxymethylcytidine
• Résultats positifs en Dot Blot: RNA,

Dilution recommandée

• Test immuno-enzymatique (ELISA): ELISA : 1:2000-1:20000
• DOT BLOT: DOT 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

Informations sur le produit

68579-1-Ig cible 5-Hydroxymethylcytidine dans les applications de Dot Blot, ELISA et montre une réactivité avec des échantillons chemical compound

• Réactivité: chemical compound
• Hôte / Isotype: Mouse / IgG1
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: PTG
• Nom complet: 5-Hydroxymethylcytidine
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine G
• Tampon de stockage: PBS with 0.02% sodium azide and 50% glycerol
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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

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