By Brian M. Ward (auth.), Barry W. Hicks (eds.)
Over the final ten years, eco-friendly Fluorescent Proteins, in addition to the opposite spectral editions, have emerged from close to obscurity to develop into a robust and flexible instrument in medical learn. In Viral purposes of eco-friendly Fluorescent Protein: equipment and Protocols, major investigators from around the globe give a contribution distinct examples of either the development and alertness of fluorescent proteins brought by means of viruses in a structure crafted to supply swift, without difficulty reproducible effects. Written within the kind of the preferred and profitable equipment in Molecular Biology™ sequence, the chapters contain short introductions to the themes, lists of the mandatory fabrics and reagents, step by step laboratory protocols, and Notes sections, which spotlight pointers on troubleshooting and fending off identified pitfalls.
Cutting-edge and straightforward to exploit, Viral functions of eco-friendly Fluorescent Protein: tools and Protocols offers researchers with an amazing advisor to the numerous makes use of of GFP and a necessary place to begin for destiny reports using this hugely adaptable protein.
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Additional info for Viral Applications of Green Fluorescent Protein: Methods and Protocols
Dejean, andP. P. Pandolfi. (2000) Role of SUMO-1-modified PML in nuclear body formation. Blood 95, 2748–2752. Chapter 4 Dual-Color Imaging of Tumor Angiogenesis Robert M. Hoffman Summary Angiogenesis is a critical step in the process of tumor metastasis. Many models have been used to study this process, but they have been artificial and do not reflect the actual process that takes place in the human being. Our laboratory has developed realistic models of angiogenesis based on orthotopic transplantation of human tumors in mice.
The development of mrfp1 has been documented elsewhere (39). The PCR-amplified mrfp1 gene fragment (without stop codon) can be inserted in the middle of the spacer listed above (at the position of Cys-Gly) to generate a flucmrfpttk triple fusion construct. 1+, is digested with HindIII and SacII and ligated in frame to PCR-amplified, HindIII/SacII-digested mrfp1 gene fragment (without stop codon) from pRSETB vector (Invitrogen) to generate the triple fusion flucmrfpttk construct (26). 2. 1. 1(+) (26).
Mix well to form a cell suspension. Pool all the cells into one 50-mL conical tube. 7. Add MEF media as needed (40-mL for one plate) for a total of ~50 mL and mix thoroughly. 8. Add 10mL of the resulting cell suspension to each of the five new 150 × 20-mm cell culture dishes. Gelatin-Coated Plates 1. 1% gelatin solution for 30min. The gelatin will solubilize. Store this solution at room temperature. 2. Place 1mL of the gelatin solution into each well of a 6-well plate, or use 6 mL gelatin solution for each 100 × 20-mm cell culture dish.
Viral Applications of Green Fluorescent Protein: Methods and Protocols by Brian M. Ward (auth.), Barry W. Hicks (eds.)