Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody, Clone RM236; 5′-hmC

$369.00

Rabbit monoclonal to 5-hydroxymethylcytosine (5-hmC), Clone RM236; 5'-hmC

Product Details
Product NameAnti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody [RM236]
Product DescriptionRabbit monoclonal to 5-hydroxymethylcytosine (5-hmC); 5'-hmC
Catalog No.31-1111-00
Clone NameRM236
Product Data SheetAnti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody Data Sheet RM236
SpecificityRM236 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.
ImmunogenBSA-conjugated 5-hydroxymethylcytosine.
ApplicationHydroxymethylated DNA immunoprecipitation (hMeDIP)
Immunohistochemistry (IHC)
Immunocytochemistry (ICC)
ELISA
Dot Blot (DB)
Flow Cytometry (FC)
Species ReactivityAll
ConjugateNone
Intended UseResearch Use Only
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Properties
FormLiquid
Storage InstructionsStore at -20.0°C
StabilityStable for 1 Year at -20.0°C from date of receipt.
Storage Buffer50% Glycerol/PBS with 1% BSA and 0.09% sodium azide
Volume/Size50 ug
Concentration 1.0 mg/mL
PurityProtein A affinity purified from an animal origin–free culture supernatant
Clone TypeMonoclonal
IsotypeRabbit IgG
UsageDot 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;
FC: 1 ug/mL - 5 ug/mL

References for 5-hydroxymethylcytosine (5-hmC) antibody [RM236]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

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)
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Product Images
ELISA

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.

Dot

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.

Immunohistochemical

Immunohistochemical staining of formalin fixed and
paraffin embedded human brain tissue sections, using
rabbit monoclonal anti-5-hmC (clone RM236) antibody.

Immunocytochemical

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.

Direct

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.

hMeDIP

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.

Flow Cytometry analysis of 5-hmC expression in HEK293 cells using Anti-5-mC rabbit monoclonal antibody [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(RM105, Red).

Flow Cytometry analysis of 5-hmC expression in HEK293
cells using Anti-5-hmC rabbit monoclonal antibody [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
(RM105, Red).



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References

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

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)