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Key Documents

MABE146

Sigma-Aldrich

Anti-5-methylcytosine Antibody, clone 33D3

clone 33D3, from mouse

Synonym(s):

methylcytosine, methylcytidine

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

33D3, monoclonal

species reactivity

human, mouse

technique(s)

Southern blotting: suitable
dot blot: suitable
flow cytometry: suitable
immunofluorescence: suitable
immunohistochemistry: suitable (paraffin)
methylated DNA immunoprecipitation (MeDIP): suitable

isotype

IgG1λ

shipped in

wet ice

target post-translational modification

unmodified

General description

5-Methylcytosine is a modified base found in the DNA of higher eukaryotes. Methylation of DNA is known to be involved in the control of gene expression and is mediated by a class of enzymes known as DNA methyltransferases (DNMT’s). Appropriate cytosine methylation is essential for normal development and maintenance of mammalian cells. Alternations in DNA methylation patterns is often associated with cancers with the turmor DNA being hypomethylated compared to DNA from normal tissues.

Specificity

Predicted to react with multiple species.
This antibody detects 5-Methylcytosine, but not unmethylated cytosine.

Immunogen

Ovalbumin-conjugated 5-methylcytosine

Application

IF
Habib, M., et al. (1999). Exp Cell Res. 249:46-53

IHC-P
Hernadez-Blazquez, F.J., et al. (2000). Gut. 47:689-693

Flow Cytometry
Reynaud, C.C., et al. (1992). Cancer Lett. 61(3):255-262

MeDIP
Keshet, I., et al. (2006). Nat Genet. 38(2):149-153
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Use Anti-5-methylcytosine Antibody, clone 33D3 (mouse monoclonal antibody) validated in DB, IF, SB, IHC(P), Flow Cytometry, MeDIP to detect 5-methylcytosine also known as methylcytosine, methylcytidine.

Quality

Evaluated by Dot Blot in methylated and unmethylated BRCA1.

Dot Blot Analysis: 1 µg/mL of this antibody detected methylcytosine in methylated and unmethylated BRCA1.

Target description

Detects methylated cytosine

Linkage

Replaces: MABE527

Physical form

Format: Purified
Protein A
Purified mouse monoclonal IgG1λ in buffer containing PBS and 0.05% sodium azide.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Analysis Note

Control
Methylated and unmethylated BRCA1

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Evidence for an instructive mechanism of de novo methylation in cancer cells.
Keshet, Ilana, et al.
Nature Genetics, 38, 149-153 (2006)
Abnormal methylation pattern in constitutive and facultative (X inactive chromosome) heterochromatin of ICF patients.
Miniou, P, et al.
Human Molecular Genetics, 3, 2093-2102 (1994)
Methylation of genomes and genes at the invertebrate-vertebrate boundary.
Tweedie, S, et al.
Molecular and cellular biology, 17, 1469-1475 (1997)
Alain Cuna et al.
American journal of respiratory cell and molecular biology, 53(1), 60-73 (2014-11-12)
DNA methylation, a major epigenetic mechanism, may regulate coordinated expression of multiple genes at specific time points during alveolar septation in lung development. The objective of this study was to identify genes regulated by methylation during normal septation in mice
Polymerase chain reaction products containing 5-methyldeoxycytidine: a microplate immunoquantitation method.
Niveleau, A, et al.
Nucleic Acids Research, 22, 5508-5509 (1994)

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