Monoclonal Anti-5-Hydroxymethylcytosine BI-HMCY

EUROPE
LIEGE SCIENCE PARK • 4102 Seraing • BELGIUM • Tel.: +32 4 372 74 00
• Fax: +32 4 372 75 00 • Toll-free: + 800 666 00 123 • [email protected]
www.eurogentec.com
NORTH AMERICA
ANASPEC - 34801 Campus Drive • Fremont, CA 94555 • USA • Tel.: +1-510-791-9560
• Toll-free: +1 877 387 6436 • Fax: +1 510-791-9572 • [email protected]
www.anaspec.com
Data Sheet
Monoclonal Anti-5-Hydroxymethylcytosine
BI-HMCY-0100
Eurogentec products are sold for research or laboratory
use only and are not to be administered to humans or
used for medical diagnostics.
Properties
Form
Liquid
DNA methylation is a major epigenetic event involved in
the control of gene expression and cell differentiation.
Maintenance of proper DNA methylation patterns is
a key process during embryo development and in the
maintenance of adult tissue homeostasis, while aberrant
DNA methylation is often associated with disease states.
5-hydroxymethylcytosine (5-hmC) is an oxidative product
of 5-methylcytosine (5-mC), catalyzed by the ten eleven
translocation (TET) family of enzymes. It is abundant in
human and mouse brains, as well as in embryonic stem
cells.
Oxidation of 5mC appears to participate in several active
DNA demethylation pathways. Hence, this event may be
important for normal processes, as well as for the global
hypomethylation which occurs during cancer development
and progression.
Storage
instructions
Store aliquoted at -20°C. Avoid
repeated freeze/thaw cycles
Storage Buffer
Phosphate Buffer Saline 10mM – NaCl
150 mM, Thimerosal 0,01 % – pH 7.4
Purity
Purified IgG fractions prepared by
affinity chromatography on Protein A
(> 95%)
Isotype
IgG1
Concentration
1 mg/ml
Validated Applications
DNA Immunoprecipitation: 1-3μg* per IP
Applications
IF: 1:500*
Description
Monoclonal antibody to
(5-hmC)
Host Species
ELISA: 1:1.000* (see fig.1)
hMeDIP:Dot
1-3µg per IP (Suggested quantity)*
Blot: 1:2.000* (see table 1)
5-Hydroxymethylcytosine
IF: 1:500 (Suggested dilution)*
(other
applications not tested)
Dot Blotting: 1:2,000 (Suggested dilution, see table 1)*
*These
values
are
quantities/dilutions. Optimal
ELISA: 1:1,000
(Suggested
dilution,
seesuggested
fig.1)*
quantity/dilution should be determined by the end user.
*Optimal quantity/dilution should be determined by the end user
Mouse
C
5-mC
5-hmC
TBS
Clone
4D9
Table 1. Dot Blot analysis of the 5hmC monoclonal antibody (4D9) with DNA standard containing Cytosine
All species
Specificity
5-hmC
Epitope
Modified 5-hydroxymethylcytosine found in DNA
vertebrates
(C), 5-methylcytosine
(5-mC)
or 5-hydroxymethylcytosine
(5-hmC).
Table
1. Dot Blot
analysis
of the 5hmC
monoclonal antibody (4D9)
of C, 5-mC and 5-hmC of the “5-Methylcytosine & 5-Hydroxymethylcytosine DNA Standard Set”
with10ng
DNA
standard containing Cytosine (C), 5-methylcytosine (5-mC) or
(D5405, Zymo Research) were spotted on a membrane (Amersham Hybond ECL).
5-hydroxymethylcytosine
(5-hmC).
The membrane was incubated with
0,5 μg/ml of the 5-hmC monoclonal antibody (dilution 1:2,000) then with
conjugated goat anti-mouse IgG (dilution 1/3000) (115-036-044, Immunotech). Specific signal
10ngperoxidase
ofonly
C, observed
5-mC
and
5-hmC
the “5-Methylcytosine & 5-Hydroxymethylcytosine
was
with 5-hmc
DNAof
standard.
DNA Standard Set” (D5405, Zymo Research) were spotted on a membrane
(Amersham Hybond ECL).
3,5
The membrane
was incubated with 0.5 μg/ml of the 5-hmC monoclonal
antibody (dilution 1:2.000) then with peroxidase conjugated goat
3,0
anti-mouse IgG (dilution 1/3.000) (115-036-044, Immunotech). Specific
signal 2,5
was only observed with 5-hmC DNA standard.
OD (450 nm)
Species Reactivity
2,0
1,5
1,0
0,5
Applications
EUROPE
LIEGE hMeDIP:
SCIENCE
•quantity)*
4102 Seraing • BELGIUM • Tel.: +32 4 372 74 00
1-3µg perPARK
IP (Suggested
IF: 1:5004(Suggested
dilution)*
• Fax: +32
372 75
00 • Toll-free: + 800 666 00 123 • [email protected]
Dot Blotting: 1:2,000 (Suggested dilution, see table 1)*
www.eurogentec.com
ELISA: 1:1,000 (Suggested dilution, see fig.1)*
*Optimal quantity/dilution should be determined by the end user
NORTH AMERICA
ANASPEC - 34801 Campus Drive • Fremont, CA 94555 • USA • Tel.: +1-510-791-9560
5-hmC
TBS
• Toll-free: +1 877 387C 6436 • 5-mC
Fax: +1 510-791-9572
• [email protected]
www.anaspec.com
Data Sheet
Table 1. Dot Blot analysis of the 5hmC monoclonal antibody (4D9) with DNA standard containing Cytosine
(C), 5-methylcytosine (5-mC) or 5-hydroxymethylcytosine (5-hmC).
10ng of C, 5-mC and 5-hmC of the “5-Methylcytosine & 5-Hydroxymethylcytosine DNA Standard Set”
(D5405, Zymo Research) were spotted on a membrane (Amersham Hybond ECL).
The membrane was incubated with 0,5 μg/ml of the 5-hmC monoclonal antibody (dilution 1:2,000) then with
peroxidase conjugated goat anti-mouse IgG (dilution 1/3000) (115-036-044, Immunotech). Specific signal
was only observed with 5-hmc DNA standard.
3,5
3,0
OD (450 nm)
2,5
2,0
1,5
1,0
0,5
0,0
100
10
1
0,1
concentration (µg/ml)
0,01
0,001
0,0001
Figure 1. Determination of the 5-hmC monoclonal antibody titer.
Figure 1. Determination of the 5-hmC monoclonal antibody titer. Direct ELISA
performed with serial dilutions of the 5-hmC monoclonal antibody (4D9) against
5-hmC in antigen coated wells. Antigen used: BSA coupled to 5-hmC base.
Estimated titer: 0.05μg/ml.
Direct ELISA performed with serial dilutions of the 5-hmC monoclonal antibody (4D9) against 5-hmC in
antigen coated wells. Antigen used: BSA coupled to 5-hmC base. Estimated titer: 0.05µg/ml.
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Related Products
Monoclonal antibody
5-methylcytosine
Starter Kit
For further information please contact our
Customer Help Desk:
For Europe:
E-mail: [email protected]
Tel: +32 4 372 76 65
BI-MECY-0100
0.1 mg
For USA:
BI-MECY-0500
0.5 mg
BI-MECY-1000
1 mg
E-mail: [email protected]
Tel.: +1-510-791-9572
BI-KHCY-0050
50 μg
Publications
Li, W., and Liu, M. (2011). Distribution of
5-hydroxymethylcytosine in different human tissues.
J Nucleic Acids 2011, 870726.
T D S - B I - H M C Y- 0 1 1 5 - V 3
Guo, Junjie U.; Su, Yijing, Zhong, Chun, Ming, Guo-li,
Song, Hongjun (2011). Hydroxylation of 5-Methylcytosine
by TET1 Promotes Active DNA Demethylation in the Adult
Brain”. Cell 145 (3): 423–434

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