ELISA of single stranded DNA using rabbit monoclonal Dot blot of double stranded DNA using rabbit monoclonal Immunohistochemical staining of formalin fixed and Immunocytochemical staining of HeLa cells using Direct ELISA of mouse brain genomic DNA using hMeDIP was performed using anti-5-hmC antibody Flow Cytometry analysis of 5-hmC expression in HEK293
Product Details
Product Name Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody, Clone RM236
Product Description Rabbit monoclonal to 5-hydroxymethylcytosine (5-hmC); 5'-hmC
Catalog No. 31-1111-00
Clone Name RM236
Product Data Sheet Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody Data Sheet RM236
Specificity RM236 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.
Immunogen BSA-conjugated 5-hydroxymethylcytosine.
Application Hydroxymethylated DNA immunoprecipitation (hMeDIP)
Immunohistochemistry (IHC)
Immunocytochemistry (ICC)
ELISA
Dot Blot (DB)
Flow Cytometry (FC)
Species Reactivity All
Conjugate None
Intended Use Research Use Only
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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 Dot Blot: 0.2 µg /mL - 1 µg /mL;
ELISA: 0.1 ug/mL - 1 ug/mL;
IHC: 0.1 µg/mL - 1 µg/mL;
ICC: 0.5 µg/mL - 2 µg/mL;
hMeDIP: 0.2 µg/mL - 2 µg/mL;
FC: 1 µg/mL - 5 µg/mL
References for 5-hydroxymethylcytosine (5-hmC) antibody [RM236] Siref AB et al. Diagnostic utility of dual 5-hmC/Melan-A immunohistochemistry in differentiating nodal nevus from metastatic melanoma: An effective first-line test for the work-up of sentinel lymph node specimen. J Cutan Pathol. (2019) 10.1111/cup.13412, IHC; Human Read more (PubMed: 30632191)
J Reichmann et al. Dual spindle formation in zygotes keeps parental genomes apart in early mammalian embryos, bioRxiv (2017) 10.1101/198275. IF; Mouse. Read more
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Product Images
anti-5-hmC (clone RM236) 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 RM236 was used as the primary antibody, and an
alkaline phosphatase conjugated anti-rabbit IgG as the
secondary antibody.
anti-5-hmC (clone RM236) 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 RM236.
paraffin embedded human brain tissue sections, using
rabbit monoclonal anti-5-hmC (clone RM236) antibody.
0.5 µg/mL anti-5-hmC antibody (RM236) (red). Actin
filaments was labeled with fluorescein phalloidin
(green). HeLa cells were fixed with 4% parafor-
maldehyde and permeabilized with methanol (−20°C)
before treatment with 2 N HCl for 30 min at 37 °C to
denature DNA.
anti-5-hmC antibody, Clone RM236. The plate was directly
coated with different concentrations of genomic DNA
isolated from mouse brain tissue. 1 µg/mL or 3 µg/mL
of RM236 was used as the primary antibody, and a
HRP conjugated anti-rabbit IgG as the secondary
antibody.
(RM236) at a 10:1 DNA:Ab ratio. 1 ng of unmethylated,
5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine
(5-hmC) DNA standard (897 bp) was spiked in 1ug 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-hmC rabbit monoclonal antibody, Clone RM236.
The cells were fixed with ice-cold MeOH, permeabilized with
0.5% Triton X-100, denatured with 2N HCl, then stained with
RM236 (anti-5-hmC, Blue) or with a negative control antibody,
Clone RM105, Red.
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