Product Details
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Product Name
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Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody [RM236]
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Product Description
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Rabbit monoclonal to 5-hydroxymethylcytosine (5-hmC); 5'-hmC
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Catalog No.
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31-1111-00
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Clone Name
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RM236
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Product Data Sheet
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Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody Data Sheet RM236
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Specificity
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RM236 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.
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Immunogen
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BSA-conjugated 5-hydroxymethylcytosine.
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Application
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Hydroxymethylated DNA immunoprecipitation (hMeDIP)
Immunohistochemistry (IHC)
Immunocytochemistry (ICC)
ELISA
Dot Blot (DB)
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Species Reactivity
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All
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Conjugate
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None
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Intended Use
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Research Use Only
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Properties
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Form
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Liquid
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Storage Instructions
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Store at -20.0°C
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Stability
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Stable for 1 Year at -20.0°C from date of receipt.
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Storage Buffer
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50% Glycerol/PBS with 1% BSA and 0.09% sodium azide
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Volume/Size
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50 ug
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Concentration
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1.0 mg/mL
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Purity
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Protein A affinity purified from an animal origin–free culture supernatant
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Clone Type
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Monoclonal
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Isotype
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Rabbit IgG
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Usage
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Dot Blot: 0.2 ug/mL – 1 ug/mL;
ELISA: 0.1 ug/mL - 1 ug/mL;
IHC: 0.1 ug/mL - 1 ug/mL;
ICC: 0.5 ug/mL- 2 ug/mL;
hMeDIP: 0.2 ug/mL- 2 ug/mL.
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References for 5-hydroxymethylcytosine (5-hmC) antibody [RM236]
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J Reichmann et al. Dual spindle formation in zygotes keeps parental genomes apart in early mammalian embryos,
bioRxiv; 10.1101/198275 (2017). IF; Mouse Read more
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.
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Product Images
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ELISA of single stranded DNA using rabbit monoclonal
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.
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Dot blot of double stranded DNA using rabbit monoclonal
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.
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Immunohistochemical staining of formalin fixed and
paraffin embedded human brain tissue sections, using
rabbit monoclonal anti-5-hmC (clone RM236) antibody.
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Immunocytochemical staining of HeLa cells using
0.5ug/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.
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Direct ELISA of mouse brain genomic DNA using
anti-5-hmC antibody (RM236). The plate was directly
coated with different concentrations of genomic DNA
isolated from mouse brain tissue. 1 ug/mL or 3 ug/mL
of RM236 was used as the primary antibody, and a
HRP conjugated anti-rabbit IgG as the secondary
antibody.
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hMeDIP was performed using anti-5-hmC 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.
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