Anti-5-methylcytosine (5-mC) Rabbit Monoclonal Antibody, Clone RM231; 5′-mC

$360.00

Rabbit monoclonal to 5-methylcytosine (5-mC) Rabbit Monoclonal Antibody, Clone RM231; 5'-mC

SKU: 31-1110-00.

Product Details
Product NameAnti-5-methylcytosine (5-mC) Rabbit Monoclonal Antibody [RM231]
Product DescriptionRabbit monoclonal to 5-methylcytosine (5-mC) Rabbit Monoclonal Antibody, 5'-mC
Catalog No.31-1110-00
Clone NameRM231
Product Data SheetAnti-5-methylcytosine (5-mC) Rabbit Monoclonal Antibody Data Sheet RM231
SpecificityRM231 reacts to 5-methylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and hydroxymethylcytosine in DNA.
ImmunogenBSA-conjugated 5-methylcytosine.
ApplicationMethylated DNA immunoprecipitation (MeDIP)
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: 0.5 ug/mL – 2 ug/mL;
ELISA: 0.1 ug/mL - 1 ug/mL;
ICC: 0.5 ug/mL - 2 ug/mL;
MeDIP: 0.2 ug/mL- 2 ug/mL;
FC: 0.5 ug/mL - 2 ug/mL.


Keywords
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Product Images
ELISA

ELISA of single stranded DNA using rabbit monoclonal
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.

Dot

Dot blot of double stranded DNA using rabbit monoclonal
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.

Immunocytochemical

Immunocytochemical staining of HeLa cells using rabbit
monoclonal anti-5-mC (clone RM231) antibody (red).
Actin filaments have been labeled with fluorescein
phalloidin (green), and nuclei stained with DAPI (blue).

Direct

Direct ELISA of HeLa cell genomic DNA using anti-5-mC
antibody (RM231). The plate was directly coated with
different concentrations of genomic DNA isolated from
HeLa cells. 1 ug/mL or 3 ug/mL of RM231 was used as
the primary antibody, and a HRP conjugated anti-rabbit IgG
as the secondary antibody.

MeDIP

MeDIP was performed using anti-5-mC 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 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-mC expression in HEK293 cells using Anti-5-mC rabbit monoclonal antibody<br />
[RM231]. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with<br />
RM231 (anti-5-mC, Blue) or with a<br />
negative control antibody (RM105, Red).

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


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