5-Hydroxymethylcytidine Monoklonaler Antikörper

5-Hydroxymethylcytidine Monoklonal Antikörper für Dot Blot, ELISA

Wirt / Isotyp

Maus / IgG1

Getestete Reaktivität

human

Anwendung

Dot Blot, ELISA

Konjugation

Unkonjugiert

CloneNo.

1E6C6

Kat-Nr. : 68579-1-Ig

Synonyme

5-Hydroxymethylcytidine, hm5C

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Galerie der Validierungsdaten

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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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Geprüfte Anwendungen

• Erfolgreiche Detektion in ELISA: 5-Hydroxymethylcytidine
• Erfolgreiche Detektion in Dot Blot: RNA

Empfohlene Verdünnung

• Enzyme-linked Immunosorbent Assay (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

Produktinformation

68579-1-Ig bindet in Dot Blot, ELISA 5-Hydroxymethylcytidine und zeigt Reaktivität mit human

• Getestete Reaktivität: human
• Wirt / Isotyp: Maus / IgG1
• Klonalität: Monoklonal
• Typ: Antikörper
• Immunogen: PTG
• Vollständiger Name: 5-Hydroxymethylcytidine
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Protein-G-Reinigung
• Lagerungspuffer: PBS mit 0.02% Natriumazid und 50% Glycerin pH 7.3.
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

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).