
5-methylcytosine (5-mC) monoclonal antibody, clone RM231

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Specifications
Product Description
Rabbit recombinant monoclonal antibody raised against 5-methylcytosine (5-mC).
Antibody Species
Rabbit
Immunogen
Original antibody is raised against 5-methylcytosine conjugated with BSA.
Sequence
N/A
Specificity
This antibody reacts to 5-methylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and hydroxymethylcytosine in DNA.
Form
Liquid
Purification
Protein A affinity purification
Isotype
IgG
Recommend Usage
Dot Blot (0.5-2 ug/mL)
ELISA (0.1-1 ug/mL)
Immunocytochemistry (0.5-2 ug/mL)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Methylated DNA Immunoprecipitation (0.2-2 ug/mL)
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS (50% glycerol, 1% BSA, 0.09% sodium azide)
Storage Instruction
Store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Immunocytochemistry
Immunocytochemistry staining of HeLa cells with 5-methylcytosine (5-mC) monoclonal antibody, clone RM231 (Cat # MAB15115) (Red). Actin filaments have been labeled with fluorescein phalloidin (Green), and nuclei stained with DAPI (Blue).Enzyme-linked Immunoabsorbent Assay
ELISA analysis of single stranded DNA with 5-methylcytosine (5-mC) monoclonal antibody, clone RM231 (Cat # MAB15115). The plate was coated with streptavidin and then biotinylated single stranded unmethylated DNA, 5-Methylcytosine (5-mC) DNA, and 5-Hydroxymethylcytosine (5-hmC) DNA. A serial dilution of MAB15115 was used as the primary antibody, and an alkaline phosphatase conjugated anti-rabbit IgG as the secondary antibody.Dot Blot
Dot blot analysis of double stranded DNA with 5-methylcytosine (5-mC) monoclonal antibody, clone RM231 (Cat # MAB15115). The membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with MAB15115.Methylated DNA Immunoprecipitation
Methylated DNA Immunoprecipitation (MeDIP) analysis of 5-methylcytosine (5-mC) monoclonal antibody, clone RM231 (Cat # MAB15115) at a 2:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1 ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input. -
Publication Reference
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Discovery of a new predominant cytosine DNA modification that is linked to gene expression in malaria parasites.
Hammam E, Ananda G, Sinha A, Scheidig-Benatar C, Bohec M, Preiser PR, Dedon PC, Scherf A, Vembar SS.
Nucleic Acids Research 2020 Jan; 48(1):184.
Application:IP, WB, Parasites, DNA, Parasites.
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Discovery of a new predominant cytosine DNA modification that is linked to gene expression in malaria parasites.
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