| Catalog Number | GTX21884 |
| Product Name | 5-Methyl Cytosine antibody |
| Product Description | Sheep polyclonal to 5-Methyl Cytosine |
| Synonyms | methyl CpG |
| Clonality | Polyclonal |
| Host | Sheep |
| Isotype | IgG |
| Immunogen | Chemical - Small Molecule conjugated to KLH. |
| Antigen Species | N/A |
| Tested Applications | IHC (Frozen sections), Immunoprecipitation, Western blot. The usefulness of this product in other applications has not been determined. |
| Application Note | WB: Use at an assay dependent dilution. IP: Use at an assay dependent dilution. This antibody can be used to precipitate enzyme-digested or sonicated genomic DNA. The use of 7 mM NaOH in the IP is crucial. ChIP: Use at a concentration of 30μg per 1μg DNA. The antibody does not work by ChIP using cross-linked chromatin but can be applied using NChIP. IHC-Fr: Use at an assay dependent dilution. Sample DNA must be denatured before use of this product so the base is unfolded and accessible to the antibody. Fixation in cold methanol for 30 minutes followed by immersion in 7 x 10-3 N NaOH for 10-15 seconds allows staining with the simultaneous detection of nuclear, cytoplasmic and membrane assigns as well as preservation of morphological detail. Not tested in other applications.concentrations should be determined by the end user. Sample DNA must be denatured before use of this product so the base is unfolded and accessible to the antibody. Fixation in cold methanol for 30 minutes followed by immersion in - x 10^-3 - NaOH for 10-15 seconds allows staining with the simultaneous detection of nuclear, cytoplasmic and membrane assigns as well as preservation of morphological detail |
| Specificity | This antibody is specific for 5-Methyl Cytosine. It was tested using immunoprecipitation against 5-Methyl Cytosine and bromodeoxyuridine (BrdU) or - control (no antigen). |
| Target | 5-Methyl Cytosine |
| Purification | Protein - purified |
| Storage Buffer | 0.15M PBS, pH7.5 |
| Storage Instruction | Aliquot and store at -20°:C long term. Avoid repeated freeze-thaw cycles |
| Notes | This antibody has also been tested in ChIP by Henk Stunnenberg but we have received feedback from customers who experienced difficulties with the antibody. We have therefore removed ChIP as an application.Henk Stunnenberg :""For optimizing ChIP assays with anti-5me-cytosine (ab1884) the following ChIP experiments were employed to analyze a known (hypermethylated) promoter:1. using the Upstate protocol on sonicated chromatin2. using the Upstate protocol on sonicated chromatin but using naked sonicated DNA3. using an adapted protocol on sonicated chromatin4. using an adapted protocol on naked, restriction enzyme-digested DNA. Throughout our tested locus, recoveries of 30-60% (ChIP / input) were obtained. Please note only the latter (4.) gave a position-dependent signal that was according the expected pattern. However, since the overall levels are rather low the antibody-to-antigen ratio most likely needs to be optimized.""" |
Image:GTX21884_2.jpgimg3122.jpgImmunoprecipitation using GTX21884 and GTX21893 (sheep anti-BrdU) vs. no Ag. Print:Print
