|
Polyclonal Antibody (Rabbit) to Human DNA Methyltransferase 1b (DNMT-1b)
Catalog Number: ABD-1B
Introduction
DNA methylation at CpG sites is an essential feature of gene regulation in higher eukarya [1-3]. Methylation of cytosine residues is correlated with silenced chromatin and there is strong evidence that inappropriate methylation can disrupt normal growth control circuits [2]. The process is mediated by enzymes called DNA methyltransferases (DNMTs) and a number of different DNMT isoforms have been identified, cloned and characterized [1]. DNMT isoforms fall into two broad groupings: 1) Maintenance activities which preferentially act on hemi-methylated DNA, and 2) De novo activites which initiate methylation on a unmethylated DNA targets. DNMT1 is a maintenance enzyme that most likely targets replicative or nascent DNA. DNMT3 type isoforms appear to be de novo methyltransferases. Other DNMTs are now known to be active but have not been classified (DNMT1b, DNMT2) while still other isoforms appear to possess key motifs of DNMT but lack sequences necessary for catalytic activity (DNMT3L). DNMT1b results from alternative splicing of the primary transcript from the Dnmt1 gene, which incorporates an in-frame insertion of 48 nt between exons 4 and 5. Moreover, the additional exon sequence is derived from the first copy of a set of seven different Alu repeats located in the fourth intron Expression of this mRNA is about 66% of the shorter Dnmt1 mRNA and this alternative splicing scheme of the Dnmt1 transcript is conserved in the higher primates (5,6). Studies by Liu et al. (4) demonstrate that DNMT1b is a fully functional and active methylase activity in humans.
Description
Anti-DNMT1b rabbit antibody was prepared using a peptide unique to the additional exon not present in DNMT-1 (5,6). The DNMT1b antibody is completely specific for the DNMT1b splice variant. Each vial contains 100 units of antibody at a concentration of 1 unit/ul. One unit corresponds to a 1:1000 dilution of stock antibody to give a working probe solution suitable for Western blotting. For example, a 1 unit/ul stock should be diluted 1:1000 to give 1 ml of probe solution.
Specificity
This antibody has been tested for use in Western blotting applications to detect the human DNMT1b. The antibody can also be used to detect human DNMT1b/DNA complexes with the ICM assay developed by Methylation, Ltd [4]. Reactivity to other species has not be examined. The antibody may be suitable for immunoprecipitation and immunohistochemistry; however, conditions optimal for these applications will have to be tested by the end user.
Storage
Store the stock antibody at -20oC for long term storage or 4oC for short term storage. The antibody is stable for 6 months from date of shipping.
Application:
This product is designed for research use only and not suitable nor approved for diagnostic procedures in animals or humans.
References
1. Robertson, K. D. (2001) Oncogene 20, 3139-355.
2. Costello, J. F., Plass , C. (2001) J Med Genet 38, 285-303.
3. Baylin, S., et al. (2001) Human Molecular Genetics 10, 687-692.
4. Liu et al., (2003) Molec. Cell. Biol. 23: 2709-2719.
5. Bonfils, C., et al. (2000) J. Biol. Chem. 275, 10754-10760.
6. Hsu, D.-W., et al. (1999) PNAS 96, 9751-9756.
SAMPLE DATA SHEET (PDF FORMAT)
|
