| 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. |
| Synonym | BRCC5; HRAD51; HsRad51; HsT16930; RAD51A; RECA |
| Protein Sequence | MAMQMQLEAN ADTSVEEESF GPQPISRLEQ CGINANDVKK LEEAGFHTVE AVAYAPKKEL
INIKGISEAK ADKILAEAAK LVPMGFTTAT EFHQRRSEII QITTGSKELD KLLQGGIETG
SITEMFGEFR TGKTQICHTL AVTCQLPIDR GGGEGKAMYI DTEGTFRPER LLAVAERYGL
SGSDVLDNVA YARAFNTDHQ TQLLYQASAM MVESRYALLI VDSATALYRT DYSGRGELSA
RQMHLARFLR MLLRLADEFG VAVVITNQVV AQVDGAAMFA ADPKKPIGGN IIAHASTTRL
YLRKGRGETR ICKIYDSPCL PEAEAMFAIN ADGVGDAKD |
| 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 |
