ChIP assays were performed using the U-2 OS (human osteosarcoma cell line), MBD1 polyclonal antibody (Cat # PAB14126) and optimized PCR primer sets. Chromatin sheared from 1x106 cells and 1.7 ug of MBD1 antibody or beads only were used per ChIP experiment. Figure shows the recovery as a % of the input DNA. Upper : Recovery by MBD1 or beads only of the MLH1 promoter, which specifically binds MBD1 (ref 1). Bottom : Recovery of the CDC6 promoter (used as a negative control) by MBD1 or beads only.
ELISA was performed using a serial dilution of MBD1 polyclonal antibody (Cat # PAB14126), crude serum and Flow Through in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 16,500.
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 (MBD). 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 (CXXC) 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. [provided by RefSeq
OTTHUMP00000163504,OTTHUMP00000163506,OTTHUMP00000163507,methyl-CpG binding domain protein 1 isoform PCM1,the regulator of fibroblast growth factor 2 (FGF-2) transcription