Human osteosarcoma cells (U-2 OS) were transfected with an expression vector for TY1-tagged MBD1. The presence of the TY1-MBD1 in the cell lysates was demonstrated by western blot analysis with the antibody directed against the TY1-tag and with MBD1 polyclonal antibody (Cat # PAB14113) (lane 2), diluted 1 : 2,000 in TBST containing 3% milk powder.
ELISA was performed using a serial dilution of MBD1 polyclonal antibody (Cat # PAB14113), 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 : 26,000.
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