ELISA of single stranded DNA using rabbit monoclonal Dot blot of double stranded DNA using rabbit monoclonal Immunocytochemical staining of HeLa cells using rabbit Direct ELISA of HeLa cell genomic DNA using anti-5-mC MeDIP was performed using anti-5-mC antibody Flow Cytometry analysis of 5-mC expression in HEK293
Product Details
Product Name Anti-5-methylcytosine (5-mC) Rabbit Monoclonal Antibody, Clone RM231
Product Description Rabbit monoclonal to 5-methylcytosine (5-mC) Rabbit Monoclonal Antibody, 5'-mC
Catalog No. 31-1110-00
Clone Name RM231
Product Data Sheet Anti-5-methylcytosine (5-mC) Rabbit Monoclonal Antibody Data Sheet RM231
Specificity RM231 reacts to 5-methylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and hydroxymethylcytosine in DNA.
Immunogen BSA-conjugated 5-methylcytosine.
Application Methylated DNA immunoprecipitation (MeDIP)
ELISA
Dot Blot (DB)
Immunocytochemistry (ICC)
Flow Cytometry (FC)
Species Reactivity All
Conjugate None
Intended Use Research Use Only
Related Product 5-Methylcytosine (5-mC) DNA Matched Antibody Pair
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Properties
Form Liquid
Storage Instructions Store at -20.0°C
Stability Stable for 1 Year at -20.0°C from date of receipt.
Storage Buffer 50% Glycerol/PBS with 1% BSA and 0.09% sodium azide
Volume/Size 50 µg
Concentration 1.0 mg/mL
Purity Protein A affinity purified from an animal origin–free culture supernatant
Clone Type Monoclonal
Isotype Rabbit IgG
Usage MeDIP: 0.2 µg/mL - 2 µg/mL;
ELISA: 0.1 µg/mL - 1 ug/mL;
Dot Blot: 0.5 µg/mL – 2 µg/mL;
ICC: 0.5 µg/mL - 2 µg/mL;
FC: 0.5 ug/mL - 2 ug/mL.
References for 5-mC antibody (RM231) Logsdon GA et al The structure, function and evolution of a complete human chromosome 8. Nature. (2021), 10.1038/s41586-021-03420-7. IF; Human. Read more (PubMed: 33828295)
Kojima N et al. Immobilization of DNA with nitrogen mustard-biotin conjugate for global epigenetic analysis. Anal Chim Acta. (2018), 10.1016/j.aca.2018.09.008. ELISA; Human. Read more (PubMed: 30392657)
Keywords
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Product Images
anti-5-mC (clone RM231) antibody. 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 RM231 was used as the primary antibody, and an
alkaline phosphatase conjugated anti-rabbit IgG as the
secondary antibody.
anti-5-mC (clone RM231) antibody. 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 RM231.
monoclonal anti-5-mC (clone RM231) antibody (red).
Actin filaments have been labeled with fluorescein
phalloidin (green), and nuclei stained with DAPI (blue).
antibody (RM231). The plate was directly coated with
different concentrations of genomic DNA isolated from
HeLa cells. 1 µg/mL or 3 µg/mL of RM231 was used as
the primary antibody, and a HRP conjugated anti-rabbit IgG
as the secondary antibody.
(RM231) 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 µg 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.
cells using Anti-5-mC rabbit monoclonal antibody, Clone RM231.
The cells were fixed with ice-cold MeOH, permeabilized with
0.5% Triton X-100, denatured with 2N HCl, then stained with
RM231 (anti-5-mC, Blue) or with a negative control antibody,
Clone RM105, Red.
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5-Methylcytosine (5-mC) DNA Matched Antibody Pair [RM231/RM476]
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Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody, Clone RM236; 5′-hmC