Purification and Quality Control The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified RAD51 is greater than 90% homogeneous based on SDS-PAGE analysis.
Unit Definition (Activity) 1 unit equals 1 nanogram of purified protein. 20 units are sufficient for a DNA-protein assay and 100 units are sufficient for a protein-protein interaction assay.
Applications RAD51 has been applied in DNA and protein-protein interactions assays.
Formulation and Storage The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles.
Background Properly controlled recombination between homologous DNA molecules (Homologous Recombination – HR) is essential for the maintenance of genome stability and for the prevention of tumorigenesis. RAD51 is a mammalian homologue of yeast RAD51 and bacterial RecA and, like its counterparts, plays a central role in HR. RAD51 coats ssDNA to form a nucleoprotein filament that invades and pairs with a homologous region in duplex DNA. It can then activate strand exchange to generate a crossover between the juxtaposed DNA molecules (1, 2). In addition to RAD51, these steps require the coordinated action of a number of other homologous-recombination proteins, including the RP-A protein, which binds single-stranded DNA, RAD52, which can bind DNA ends, anneal complementary single-stranded DNA molecules and enhance the specificity of RAD51, and a number of RAD51 paralogs (3). The tumour-suppressor proteins BRCA1 and BRCA2 colocalize with RAD51 in DNA-damage-induced nuclear foci (4). BRCA2 has been shown to interact directly with RAD51 (5) and thus plays a direct role in repair by HR, through control of the availability and function of the central mediator, RAD51. Recombinant His tagged RAD51 was expressed in a strain of E. coli and purified by affinity and FPLC chromatography.
References: 1. Modesti M. et al., (2001) Genome Biol. 2(5) Epub 2. Ashok R Venkitaraman, (2001) Curr Op Cell Biol. 13, 338-343 3. Hiom K., (2000) Curr Biol 9, 446-448 4. West, SC, (2003) Nat Rev Mol Cell Biol.6, 435-445. 5. A.A. Davies et al., (2001) Mol Cell 7, 273–282
DISCLAIMER
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
