| Species : |
Human |
| Source : |
Bacteria |
| Tag : |
His |
| Description : |
This is an N-ras oncogene encoding a membrane protein that shuttles between the Golgi apparatus and the plasma membrane. This shuttling is regulated through palmitoylation and depalmitoylation by the ZDHHC9-GOLGA7 complex. The encoded protein, which has intrinsic GTPase activity, is activated by a guanine nucleotide-exchange factor and inactivated by a GTPase activating protein. Mutations in this gene have been associated with somatic rectal cancer, follicular thyroid cancer, autoimmune lymphoproliferative syndrome, Noonan syndrome, and juvenile myelomonocytic leukemia. |
| Form : |
White lyophilized powder |
| Bio-activity : |
The biological activity of N-Ras can be determined from the ability of the RasGRF1 exchange domain (Ras-GRF1-ExD) to catalyze the exchange of GDP for GTP on N-Ras. A standard biological assay for monitoring the biological activity of N-Ras is an exchange assay utilizing the 2× Exchange Buffer from the RhoGEF exchange assay kit and the human RasGRF GEF domain. |
| Molecular Mass : |
25 kDa |
| Purity : |
Protein purity is determined by scanning densitometry of Coomassie Blue-stained protein on a 4-20% polyacrylamide gradient gel. His tagged N-Ras protein was determined to be > 90% pure. |
| Applications : |
Study of N-Ras exchange activity with different GEFs, Identification of N-Ras exchange factors (GEFs), Positive control for GEF studies, Biochemical characterization of N-Ras protein interactions, Western blot standard |
| Stability : |
The protein is stable for six months if stored at -70 centigrade. The protein should not be exposed to repeated freezethaw cycles. The lyophilized protein is stable at 4 centigrade desiccated (<10% humidity) for one year. |
| Storage : |
In order to maintain high biological activity of the protein, it is strongly recommended that the protein solution be supplemented with DTT to 1 mM final concentration, aliquoted into "experiment-sized" amounts, snap frozen in liquid nitrogen, and stored at -70 centigrade. |
| Reconstitution : |
Before reconstitution, briefly centrifuge to collect the product at the bottom of the tube. The protein should be reconstituted to 5 mg/mL with the addition of 20 μL of Milli-Q water (100 μg size). When reconstituted, the protein will be in the following buffer: 50 mM Tris pH 7.5, 50 mM NaCl, 0.5 mM MgCl2, 5% (w/v) sucrose, and 1% (w/v) dextran. |
