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| Western Blot | Chromatin IP | Immunoprecipitation |
|---|---|---|
| Not tested. | Not tested. | 1 µg per IP |
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Background
5-Methylcytosine (5-Methylcytidine) is a modified base that is found in the DNA of plants and vertebrates. DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. The 5-methylcytidine antibody (Clone 33D3) has been developed to discriminate between the modified base and its normal cytosine counterpart, allowing for gene promoter methylation analysis.
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Immunogen
Clone 33D3 recognizes the modified base 5-methylcytidine found in plant and vertebrate DNA.
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Application Notes
This 5-Methylcytosine antibody has been validated for use in applications including flow cytometry (1:500), dot blot (1:1,000) immunohistochemistry (1:500) and methylated DNA immunoprecipitation (MeDIP, 1 µg per IP).
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Buffer
Purified IgG in 10 mM phosphate, 0.15 M NaCl and 0.01% thimerosal.
Bridging Antibody to Improve ChIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP and IP experiments that use 5-Methylcytosine (5-mC) antibody (mAb).
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)References
Reynaud, C. and Nivelleau A. (1990) Annal. Letters 23: 31-45. Sasco, A.J. et. al. (1996) Cancer Letters 108: 157-162. Rougier, N. et. al. (1998) Genes and development 14: 2108-2113. Habib, M. et. al. (1999) Exp. Cell Res. 249: 46-53
MeDIP assay using 5-Methylcytosine mAb (Clone 33D3). Human DNA (500 ng) digested with MseI was subjected to MeDIP and then analyzed by quantitative real-time PCR with primers specific for genes that are normally methylated (Xist, NYESO, and SNRPN) or unmethylated (APC, p48). The MeDIP'd DNA (enriched DNA) was plotted.
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for 5-Methylcytosine (5-mC) antibody (mAb) (Clone 33D3)Guarantee
This product is guaranteed for 6 months from date of arrival.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
