Bioluminescence: Methods and Protocols by Sung Bae Kim

By Sung Bae Kim

This quantity offers precise laboratory protocols concerning the 3 significant course applied sciences luciferases, effective optical probes, and purposes to visualizing molecular occasions in dwelling subjects. Bioluminescence: equipment and Protocols, 3rd Edition guides readers via chapters on materials of bioluminescent probes, and fabrication of bioluminescent probes, functions to dwelling matters and instrumentations. Written within the hugely winning Methods in Molecular Biology series layout, chapters comprise introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and pointers on troubleshooting and fending off recognized pitfalls.

Authoritative and functional, Bioluminescence: tools and Protocols, 3rd Edition is an invaluable supplement to the 1st and moment version for brand spanking new and skilled researchers alike.

 

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To efficiently prepare the organozinc reagent, zinc chloride needs to be sufficiently dried before addition of benzylmagnesium chloride. 3. Benzylmagnesium chloride should be used and stored under inert gas. We find that it is best to prepare this fresh. 4. 3,5-Dibromopyrazin-2-amine (1) needs to be dried enough before adding. 30 Ryo Nishihara et al. 5. Tetrakis(triphenylphosphine)palladium(0) is unstable under aerobic conditions. The reaction mixture needs to be deaerated before and after adding of palladium catalyst.

1. Native CTZ (nCTZ, Biotium) (store at −30 °C) (see Note 1). 2. DeepBlueC™ (Biotium) (store at −30 °C). 3 Components for Bioluminescence Assay 21 1. 75) (Promega, Madison, WI, USA). 2. ). 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.

Ikeo, 2014-A63, 2015-A47). References 1. Verhaegen M, Christopoulos T (2002) Recombinant Gaussia luciferase. Overexpression, purification, and analytical application of a bioluminescent reporter for DNA hybridization. Anal Chem 74:4378–4385 2. Markova SV, Golz S, Frank LA, Kalthof B, Vysotski ES (2004) Cloning and expression of cDNA for a luciferase from the marine copepod Metridia longa. A novel secreted bioluminescent reporter enzyme. J Biol Chem 279:3212–3217 3. Takenaka Y, Masuda H, Yamaguchi A, Nishikawa S, Shigeri Y, Yoshida Y, Mizuno H (2008) Two forms of secreted and thermostable luciferases from the marine copepod crustacean, Metridia pacifica.

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