By Sung Bae Kim
This quantity provides specific laboratory protocols in regards to the 3 significant path applied sciences luciferases, effective optical probes, and functions to visualizing molecular occasions in dwelling subjects. Bioluminescence: tools and Protocols, 3rd Edition guides readers via chapters on elements of bioluminescent probes, and fabrication of bioluminescent probes, functions to dwelling topics and instrumentations. Written within the hugely profitable Methods in Molecular Biology series structure, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and pointers on troubleshooting and fending off recognized pitfalls.
Authoritative and useful, Bioluminescence: tools and Protocols, 3rd Edition is an invaluable supplement to the 1st and moment variation for brand spanking new and skilled researchers alike.
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Additional resources for Bioluminescence: Methods and Protocols
2 Materials Used buffers 1. Phosphate-buffered saline (PBS) 2. Hank’s balanced salt solution buffer (HBSS, Gibco) 3. Lysis buffer (E291A; Promega) included in a bioluminescence assay kit (E2820, Promega) 4. Renilla luciferase (RLuc) assay buffer (E290B, Promega) included in a bioluminescence assay kit (E2820, Promega) Cells and protein expression-related reagents 5. 1(+) vector (Invitrogen) for mammalian cell expression 6. pColdIV vector (Takara) for E. coli expression 7. 1 vector (Toyobo) for mammalian cell expression 8.
6 Synthesis of (E)-4-(2-(5-Amino-6benzylpyrazin-2-yl)vinyl) phenol (See Fig. 1 Compound ( 6)) Yield 24 % (yellow solid compound). Eluent composition: n-hexane/ethyl acetate = 80/20 to 50/50. 16 (s, 2H). 8. 1814 [M + H]+. 1. ) in dichloromethane (12 mL) (see Note 6). 2. 0 M boron tribromide dichloromethane solution (12 mL) slowly into the solution at 0 °C, followed by stirring for 13 h at 40 °C (see Note 7). 3. After cooling to room temperature, add saturated NaHCO3 aq. into the reaction mixture to neutralize, and evaporate to remove most of the solvent.
3. Native CTZ (nCTZ, NanoLight Technologies, Pinetop, AZ, USA). 4. TransIT-LT1 transfection reagent (Takara, Osaka, Japan). 5. Lysis buffer (E291A) (Promega, Madison, WI, USA). 6. Hanks’ balanced salt solution (HBSS). 1 General Procedure for Synthesis 1. Carry out all moisture-sensitive reactions under an atmosphere of argon. 2. The composition of mixed solvents is given by the volume ratio (v/v). 3. ) spectrometer at room temperature. 4. The measurement for 1H-NMR is performed at 500 MHz. 5. The measurement of 13C-NMR is performed at 125 MHz or 150 MHz.