B : Western blot was performed on nuclear extracts from the U-937 (human leukemic monocyte lymphoma cell line ; 40 ug) with MBD4 polyclonal antibody (Cat # PAB14057), diluted 1 : 2,000 in TBST containing 3% milk powder.
Western Blot (Transfected lysate)
A : Human osteosarcoma cells (U-2 OS) were transfected with an expression vector for TY1-tagged MBD4. The presence of TY1-MBD4 in the cell lysates was demonstrated by western blot analysis with the antibody directed against the TY1-tag (Lane 1) and with the MBD4 polyclonal antibody (Cat # PAB14057) (Lane 2), diluted 1 : 2,000 in TBST containing 3% milk powder.
ELISA was performed using a serial dilution of MBD4 polyclonal antibody (Cat # PAB14057) in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 15,000.
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. MBD4 may function to mediate the biological consequences of the methylation signal. In addition, MBD4 has protein sequence similarity to bacterial DNA repair enzymes and thus may have some function in DNA repair. Further, MBD4 gene mutations are detected in tumors with primary microsatellite-instability (MSI), a form of genomic instability associated with defective DNA mismatch repair, and MBD4 gene meets 4 of 5 criteria of a bona fide MIS target gene. [provided by RefSeq
3,N(4)-ethenocytosine glycosylase,G/5-fluorouracil mismatch glycosylase with biphasic kinetics,G/T mismatch glycosylase,G/U mismatch glycosylase,putative methyl-CpG binding protein