Anticorps Monoclonal anti-5-methylcytosine

5-methylcytosine Monoclonal Antibody for IHC, Dot Blot, ELISA

Hôte / Isotype

Mouse / IgG2b

Réactivité testée

m5C et plus (2)

Applications

WB, IHC, IF, RIP, Dot Blot, ELISA

Conjugaison

Non conjugué

Publications(9)

CloneNo.

2C9G9

Avis (2)

N° de cat : 68301-1-Ig

Synonymes

m5C, 5mC

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Galerie de données de validation

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  IHC staining of mouse testis using 68301-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68301-1-Ig (5-methylcytosine antibody) at dilution of 1:5000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse testis using 68301-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68301-1-Ig (5-methylcytosine antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 68301-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 68301-1-Ig (5-methylcytosine antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat testis using 68301-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat testis tissue slide using 68301-1-Ig (5-methylcytosine antibody) at dilution of 1:5000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  ELISA experiment of compound using 68301-1-Ig

  Indirect ELISA and competitive ELISA results show that this antibody is specific to m5C. Indirect ELISA (blue curve, refer to top X-right Y axis) was performed by coating BSA conjugated m5C at 0.4ng/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 m5C or its structure analogue compound Cytidine was mixed in 20ng/mL primary antibody (red curve, refer to bottom X-left Y axis).

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  Dot Blot experiment of HeLa using 68301-1-Ig

  Total DNA 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-methylcytosine antibody 68301-1-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 IgG2b isotype control Monoclonal antibody 66360-3-Ig at the same dose.

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Applications testées

• Résultats positifs en IHC: tissu testiculaire de souris, tissu cérébral de rat, tissu testiculaire de rat; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en ELISA: compound,
• Résultats positifs en Dot Blot: cellules HeLa,

Dilution recommandée

• Immunohistochimie (IHC): IHC : 1:2500-1:10000
• Test immuno-enzymatique (ELISA): ELISA : 1:10-1:100
• DOT BLOT: DOT BLOT : 1:2500-1:10000

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

Applications publiées

• WB: See 1 publications below
• IHC: See 1 publications below
• IF: See 1 publications below
• RIP: See 1 publications below

Informations sur le produit

68301-1-Ig cible 5-methylcytosine dans les applications de WB, IHC, IF, RIP, Dot Blot, ELISA et montre une réactivité avec des échantillons m5C

• Réactivité: m5C
• Réactivité citée: Humain, souris
• Hôte / Isotype: Mouse / IgG2b
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: Composé
• Nom complet: 5-methylcytosine
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine A
• 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

In eukaryotes, 5-methylcytosine in DNA (5mC) and its oxidized derivatives (5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC)) are the most prominent modifications and have been suggested to contribute to epigenetic gene regulation through a variety of different mechanisms. m5C has also been found in mRNA, rRNA and tRNA of representative organisms from all kinds of species. As reversible epigenetic modifications, m5C modifications of RNA affect the fate of the modified RNA molecules and play important roles in various biological processes including RNA stability control, protein synthesis, and transcriptional regulation (PMID: 35365216). This antibody can recognize both 5mC in DNA and m5C in RNA.

Protocol for Dot Blot:
https://www.ptglab.com/protocol/68301-1-IgDotBlot.pdf

Publications

Species	Application	Title
human		Cancer Lett; NSUN4 mediated RNA 5-methylcytosine promotes the malignant progression of glioma through improving the CDC42 mRNA stabilization; Authors - Zhen Zhao
human		Biomedicines; NSUN2 Promotes Head and Neck Squamous Cell Carcinoma Progression by Targeting EMT-Related Gene LAMC2 in an m5C-YBX1-Dependent Manner; Authors - Shuojin Huang
human	WB	Redox Biol; NSUN2 lactylation drives cancer cell resistance to ferroptosis through enhancing GCLC-dependent glutathione synthesis; Authors - Kaifeng Niu
		Cancer Immunol Immunother; NSUN2/ALYREF axis-driven m5C methylation enhances PD-L1 expression and facilitates immune evasion in non-small-cell lung cancer; Authors - Yiran Yang
human	IHC	J Exp Clin Cancer Res; NONO regulates m5C modification and alternative splicing of PTEN mRNAs to drive gastric cancer progression; Authors - Gaichao Zhao
		NPJ Precis Oncol; piRNA28846 has the potential to be a novel RNA nucleic acid drug for ovarian cancer; Authors - Hong-Lei Qin

Avis

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Lisa (Verified Customer) (08-20-2025)

I used this Antibody for slot-blotted RNA and it worked well with low background.

• Applications: Western Blot, Other
• Primary Antibody Dilution: 1:5000

FH

DONGHONG (Verified Customer) (07-02-2024)

I used this antibody for the competitive elisa kit. Coating BSA-m5C and blocking the 96-well plate. Use the the standard to competitive bind m5C antibody. However, the standard curve was not good as m5C from Abcam (ab10805).

• Applications: Other
• Primary Antibody Dilution: 1:1000

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