Western Blot: Detects a band of approximately 120 kDa (predicted molecular weight: 102 kDa). Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min. Use in chromatin immunoprecipitation reported in scientific literature (PMID 24623306)
Methylation of DNA at cytosine residues plays an important role in regulation of gene expression, genomic imprinting and is essential for mammalian development. Hypermethylation of CpG islands in tumor suppressor genes or hypomethylation of bulk genomic DNA may be linked with development of cancer. To date, 3 families of mammalian DNA methyltransferase genes have been identified which include Dnmt1, Dnmt2 and Dnmt3. Dnmt1 is constitutively expressed in proliferating cells and inactivation of this gene causes global demethylation of genomic DNA and embryonic lethality. Dnmt2 is expressed at low levels in adult tissues and its inactivation does not affect DNA methylation or maintenance of methylation. The Dnmt3 family members, Dnmt3a and Dnmt3b, are strongly expressed in ES cells but their expression is down regulated in differentiating ES cells and is low in adult somatic tissue. Recently, it has been shown that naturally occurring mutations of Dnmt3b gene occurs in patients with a rare autosomal recessive disorder, termed ICF (immunodeficiency, centromeric instability, and facial anomalies) syndrome.
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FAQs for DNMT3A (3)
Do you have any information about the epitope of your antibody, so I could check the chances for the antibody working in the honey bee?
This antibody was raised against bacteria expressed recombinant mouse Dnmt3a. The epitope was found to lie near the C-terminus (a.a. 705-908), see Chen et (2002) for details. We haven't mapped the binding binding epitope although Chen et (2002) mapped it to a range between 705-908 amino acids: I am not sure what technique you are planning with the antibody, but if you are doing a western blot, we do have a western blot postive control ready to use lysate NBP2-25063 and I would l highly suggest using that.
In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
I was wondering what does Format 7C mean as far as anitbodies go?
7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
Go Ahead! Make My DNA DNA methylation plays a critical role the long-term silencing of transcription and is essential for processes such as embryonic development, germline differentiation, and tissue maturation. Dnmt3a is a member of the C5-methyltransferase family that re... Read full blog post.