From tissues, extraction

A 17 amino acid long peptide sequentially related to opioid peptides in particular dynorphin A. OFQ/N is inactive at the 5, k, and p opioid receptors, but binds to its own NOP receptor (formerly ORL-1, for opioid receptor like-1). In contrast to opioid peptides, OFQ/N has no direct analgesic properties. OFQ/N is the first example for the discovery of a novel neurotransmitter from tissue extracts by using an orphan receptor as bait. Centrally administered in rodents, OFQ/N exerts anxiolytic properties. OFQ/N agonists and antagonists... 

Another similar technique to Southern blotting is Northern blotting. Here, instead of DNA fragments, mRNA fragments are probed with a labelled cDNA probe after separation by electrophoresis and transfer to nitrocellulose membranes. Northern blotting is used to detect and quantify mRNA from tissue extracts. 

Saelens et al. measured choline, acetylcholine, and their metabolites by combined enzymatic and radiometric techniques [219]. Choline and acetylcholine were isolated from tissue extracts by a low voltage... 

IEF was first successfully applied to proteins in 1938, when it was used to separate the protein hormones vasopressin and oxytocin from tissue extracts. Twenty years later, ampholytes were first focused in a continuous pH gradient, stabilized by a dense sucrose medium, as an alternative to the multicompartment method. The continuous pH gradient in the sucrose medium was established by allowing acid and base to diffuse into opposite ends of the sucrose medium, held in a U-cell, from their respective electrode chambers. The stabilization of this continuous pH gradient with carrier ampholyte species led to modern IEF methods. 

In standard MS proteomics, well-established and efficient protocols allow for the en masse identification of proteins from tissue extracts. A key step within this process is the initial separation and purification of the molecules before submitting them to the mass spectrometer. However, in MSI studies, a general identification strategy for proteins, peptides, and metabolites is still missing due to several reasons. First, in comparison to LC-MS-based... 

Sugar phosphates and nucleotides from tissue extracts Anion exchange chromatography orcinol-H2SO4 detection Blanshard [245]... 

One final word of advice for the potential chromatographer who intends to separate and quantitate phospholipids from tissue extracts using HPLC with any of the chromatographic columns described earlier is to use a guard column to enhance the lifetime of the analytical column. 

Fat from tissue extracted with solvent cleaned up by column chromtography and concentrated... 

The fluorometric and chromatographic techniques described in this chapter can be valuable in all phases of peptide chemistry and biochemistry. For quantitative analysis, they can provide a degree of sensitivity and specificity not always attainable with bioassay or immunoassay. They allow preparative isolations of peptides from tissue extracts to be carried out at a level that is orders of magnitude lower than is possible by classical means. They... 

A number of recent papers deal with Immunoassay techniques, especially various enzyme-linked immunosorbent assays (ELISA). Detection limits for HPLC and ELISA are quite comparable, but the ELISA technique suffers from falsepositive results, because of cross-reactions, and, more importantly, false-negative results, especially from tissue extracts (Monaci Palmisano, 2004). 

We describe here methods for analyzing the interaction of fragments of Tuba with lipids and with dynamin. The property of the DH-BAR module of Tuba to bind liposomes in a cosedimentation essay (Fig. 1C) supports the hypothesis that the BAR domain of Tuba localizes and/or activates its DH domain at the membrane. The juxtaposition of four SH3 domains that bind dynamin generates a module with a striking avidity for this GTPase. Thus, the NH2-terminus of Tuba is a very convenient tool to deplete dynamin from tissue extracts as well as to affinity-purify this protein from tissue and cell extracts to near purity in one step. 

The adsorption of alkaloids from tissue extracts by suitable adsorbents had been considered by Daubney and Nickolls, but they found that quantitative results were not obtained in the presence of strong ammonium sulphate solutions, the use of which is a necessary part of their extraction process. 

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