- Antibody Type: Monoclonal
- Isotype: IgG
- Purification: Protein A Chromatography
- Host: Mouse
- Reactivity: Human, Not Species Specific
- MeDIP: 1 µg per IP
- IHC (FFPE): 1:1000
- WB: 1:250 dilution
- ELISA: 1:10,000 dilution
注意：Methods for detecting 5-mC in Immunocytochemistry may be found in the following publication: Inoue, A. and Zhang, Y. 2011. Science. Oct 14;334(6053):194. doi: 10.1126/science.1212483. Epub 2011 Sep 22.
Clone 33D3 recognizes the modified base 5-methylcytidine found in plant and vertebrate DNA.
Purified IgG in 10 mM phosphate, 0.15 M NaCl and 0.01% thimerosal. Thimerosal is highly toxic.
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.
Active Motif's Human Positive Control Primer Set ZC3H13 (Cat. No. 71009) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Negative Control Primer Set 1 (Cat. No. 71011) has been validated for both qPCR and endpoint PCR.
Bridging Antibody to Improve ChIP, MeDIP 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, MeDIP 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, MeDIP and IP experiments that use 5-Methylcytosine (5-mC) antibody (mAb).
Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with 5-Methylcytosine (5-mC) antibody (mAb).
Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
ChIP = Chromatin Immunoprecipitation; DB = Dot Blot; ELISA = Enzyme-linked Immunosorbent Assay;EMSA = Electrophoretic Mobility Shift Assay; FC = Flow Cytometry; ICC = Immunocytochemistry; IF = Immunofluorescence; IHC = Immunohistochemistry; IP = Immunoprecipitation; MeDIP = Methyl-DNA Immunoprecipitation; TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer; WB = Western Blotting
5-Methylcytosine (5-mC) antibody (mAb)