ChIP assays were performed using human osteosarcoma (U-2 OS) cells, the anti-MBD1 antibody (Cat. no. 49-1027), and optimized PCR primer sets. Each ChIP assay used sheared chromatin from 1 million cells and 1.7 µg of MBD1 antibody or beads only. This figure shows the recovery as a percentage of the input DNA. Left: Recovery by MBD1 or beads only of the MLH1 promoter, which specifically binds MBD1. Right: Recovery of the CDC6 promoter (used as a negative control) by MBD1 or beads only.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Raised against the human MBD1 protein.|
|Contains||<0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1.7 ug|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
DNA methylation is the major modification of eukaryotic genomes and plays an essential role in mammalian development. Human proteins MECP2, MBD1, MBD2, MBD3, and MBD4 comprise a family of nuclear proteins related by the presence in each of a methyl-CpG binding domain. Each of these proteins, with the exception of MBD3, is capable of binding specifically to methylated DNA. MECP2, MBD1 and MBD2 can also repress transcription from methylated gene promoters. Five transcript variants of the MBD1 are generated by alternative splicing resulting in protein isoforms that contain one MBD domain, two to three cysteine-rich domains, and some differences in the COOH terminus. All five transcript variants repress transcription from methylated promoters; in addition, variants with three CXXC domains also repress unmethylated promoter activity. MBD1 and MBD2 map very close to each other on chromosome 18q21.
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