Zhizhong Z. Pan's Opioid Research: Methods and Protocols (Methods in Molecular PDF

By Zhizhong Z. Pan

Opioid study is without doubt one of the multidisciplinary examine parts that contain complicated recommendations starting from molecular genetics to neuropharmacology, and from behavioral neuroscience to medical medication. In present opioid examine, it has develop into more and more vital to take advantage of a number of ways at molecular, mobile, and process degrees for investigations on a selected opio- comparable objective approach. that regularly calls for realizing and utilising cro- box strategies and techniques for the good fortune of one’s examine initiatives. via its wide spectrum of assurance, Opioid learn: equipment and Protocols presents a complete number of significant laboratory equipment and protocols in present opioid examine, protecting issues from molecular and genetic strategies to behavioral analyses of animal types, after which to scientific perform. it's going to function a handy reference publication from which these inquisitive about opioid examine will examine or excellent the required cross-field thoughts. The distinct tools and protocols defined in Opioid examine: tools and Protocols have every one been effectively utilized in present opioid study. half I presents molecular concepts for the cloning and expression of opioid receptors, and for the quantitative characterization in their signaling pathways. half II contains fundamental thoughts for mapping the distributions and detecting the expression degrees of opioid receptors, opioid peptides, and their messages in mind tissues and in person cells. half III bargains with tools for growing in vitro receptor versions and in vivo animal versions to check opioid capabilities. half IV describes sensible purposes of opioids in medical drugs for the remedy of ache and opioid dependancy.

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The reaction time should be kept to a minimum. 6. The assay is based on the changes in the net charge of the PKA substrates before and after phosphorylation. Phosphorylation by PKA of PKA specific substrate (LRRASLG) alters the peptide’s net charge from positve to negative. This allows the separation of phosphorylated and nonphosphorylated substrate by electrophoresis on an agarose gel at neutral pH. The phosphorylated species migrates toward the positive electrode (the same direction as DNA moves during electrophoresis) while the nonphosphorylated substrate migrates toward the negative electrode (opposite to the direction DNA moves).

3. , Hurley, J. , and Yu, L. (1993) Molecular cloning and functional expression of a µ-opioid receptor from rat brain. Mol. Pharmacol. 44, 8–12. 4. Wang, J. , Eppler, C. , Spivak, C. , and Uhl, G. R. (1993) µ opiate receptor: cDNA cloning and expression. Proceed. Nat. Acad. Sci. USA 90, 10,230–10,234. Expression of Opioid Receptors 27 5. Thompson, R. , and Watson, S. J. (1993) Cloning and pharmacological characterization of a rat µ opioid receptor. Neuron 11, 903–913. 6. , et al. (1993) Cloning and expression of a cDNA for the rat kappa-opioid receptor.

DMSO shock can increase transfection efficiency by 2–3 folds in our CHO cells, but it may not be necessary for other cell lines. The shock time, from 90–120 s, but no more than 120 s, should be followed to avoid overshocking the cells. 10. Many types of liposome-mediated transfection reagents are available from same or different companies. There are also several formulas even with the same type of lipid. For instance, LipofectAmine has three different formulas, LipofectAmine 2000, LipofectAmine Plus and LipofectAmine.

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