| ALX-201-261-C010 | 10 µg | 401.00 USD |  |
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| Alternative Name: | nNOS, NOS I |
| MW: | ~160kDa/subunit; homodimer. |
| Source: | Produced in E. coli. |
| EC: | 1.14.13.39 |
| UniProt ID: | P29475 |
| Concentration: | ~1mg/ml |
| Formulation: | Liquid. In 40mM bis-tris-propane, pH 7.8, containing 150mM sodium chloride, 0.1% BSA, 5µM tetrahydrobiopterin, 0.5mM DTT and 5% glycerol. |
| Purity: | ≥90% (SDS-PAGE and heme spectral analysis) |
| Specific Activity: | ≥400-600nmol/min/mg (U/mg) protein. Amount assayed 0.5-1µg. Optimal conditions: 37°C, 40mM TRIS-HCl, HEPES or EPPS buffer, pH 7.6, 10µM oxyhemoglobin, 0.3mM DTT, 1mM NADPH, 4µM FAD, 4µM FMN, 4µM H4B, 100units/ml catalase, 10units/ml superoxide dismutase, 0.1mg/ml BSA, 35µl of 9mM EDTA, 35µl of 150mg/ml calmodulin and 12.5mM CaCl2 in a final volume of 1ml. |
| Shipping: | Shipped on Dry Ice |
| Long Term Storage: | -80°C |
| Use/Stability: | Stable for at least 1 year after receipt when stored at -80°C. Keep stock vial on ice at all times! |
| Handling: | Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C. |
| Protocol: | Preparation of oxyhemoglobin: 1) Wash a G-50 column with your assay buffer of your choice. 2) Weigh out 2-3mg hemoglobin in eppendorf tube and add 200µl of your column buffer. 3) Add several crystals of dithionite to reduce hemoglobin to oxyhemoglobin. 4) Drip onto G-50 column. 5) Wash with 500µl buffer; repeat a couple of times. 6) Collect brownish eluate in eppendorf tube and store on ice. 7) Wash column with water and reuse again. 8) Use ~10-15µl of oxyhemoglobin in the assay corresponding to ~10µM. |
| Regulatory Status: | RUO - Research Use Only |
