DNA methylation, or the addition of methyl groups to cytosine bases in the dinucleotide CpG, is imperative to proper development and regulates gene expression.
The methylation pattern involves the enzymatic processes of methylation and demethylation.
The demethylation enzyme was recently found to be a mammalian protein, which exhibits demethylase activity associated to a methyl-CpG-binding domain (MBD) (1).
The enzyme is able to revert methylated cytosine bases to cytosines within the particular dinucleotide sequence mdCpdG by catalyzing the cleaving of the methyl group as methanol.
MeCP2 and MBD1 (PCM1) are first found to repress transcription by binding specifically to methylated DNA (2).
MBD2 and MBD4 (also known as MED1) were later found to colocalize with foci of heavily methylated satellite DNA and believed to mediate the biological functions of the methylation signal.
Surprisingly, MBD3 does not bind methylated DNA both in vivo and in vitro.
MBD1, MBD2, MBD3, and MBD4 are found to be expressed in somatic tissues, but the expression of MBD1 and MBD2 is reduced or absent in embryonic stem cells, which are known to be deficient in MeCP1 activity.
MBD4 have homology to bacterial base excision repair DNA N-glycosylases/lyases (3).
In some microsatellite unstable tumors MBD4 is mutated at an exonic polynucleotide tract (4).
Applications:Can be used for detection of MBD1 (isoform 4, a 503 amino acid protein) by Western blot.
Western blot analysis of MBD1 in HeLa cell lysate with anti-MBD1 pcAb.
A protein band with an approximate molecular weight of 55kD was detected.
Recommended Dilution:Western blot: 2ug/ml
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are stable for at least 12 months at -20°C.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
仕様
Size：100ug
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein G affinity chromatography.
Form：Supplied as a liquid in PBS, 0.02% sodium azide.
Specificity：Recognizes human MBD1 (Methyl CpG Binding Protein 1)
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Hu
Immunogen：Synthetic peptides corresponding to amino acids 98-113 and 342-356 of human MBD1.
