Advanced Methods in Protein Microsequence Analysis by Brigitte Wittmann-Liebold, Johann Salnikow

By Brigitte Wittmann-Liebold, Johann Salnikow

Much of the hot brilliant development within the organic sciences might be at­ tributed ot the facility to isolate, examine, and structurally represent proteins and peptides that are found in cells and mobile organelles in just very small quantities. fresh advances in protein chemistry and particularly the applying of latest micromethods have ended in fruitful advances within the knowing of easy mobile procedures. components the place protein-chemical experiences have ended in curiosity­ ing discoveries comprise the peptide hormones and their unencumber components, development components and oncogenes, bioenergetics, proton pumps and ion pumps and chan­ nels, topogenesis and protein secretion, molecular virology and immunology, membrane protein research, and receptor study. in reality, the major equipment are actually to be had to resolve a few of the significant awesome difficulties of molecular biology and particularly questions of basic curiosity which relate to devel­ opmental biology and specificity in cell-cell interplay. during this quantity now we have assembled descriptions of techniques that have re­ cently been proven to be efficaceous for the isolation, purification, and chemical characterization of proteins and peptides which are basically on hand in minute quantities. Emphasis is put on well-established micromethods that have been established and located invaluable in lots of laboratories by way of skilled investigators. The chapters are written by means of experts, and describe quite a number delicate suggestions which are utilized by researchers operating in laboratories with in simple terms modest assets and equipment.

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4 ml acetic acid 1 ml5M KOH add bidistilled water to 90 ml, dissolve, fill up to 100 ml with bidistilled water, filter, and store at 4°C. 0. -J. Brockmoller and R. M. 5 g agarose, boil until solution is completely clear and use at about 50°C. 5 1 methanol 375 ml acetic acid fill up to 51 with bidistilled water, dissolve, filter. 10 Destaining Solution 1 1 acetic acid 11 methanol fill up to 101 with distilled water. All reagents used were pro analysis grade, purchased from Merck; acrylamide (2 X crystallized) and bis-acrylamide (2 X crystallized) were purchased from Serva.

HPLC Quantitative Analysis of DABS-Amino Acids ............................. Comments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56 57 57 57 57 57 58 59 59 60 DABS-Cl = dimethylaminoazobenzene sulfonyl chloride; DABS = dimethylaminoazobenzene sulfonyl; DNS-Cl = dimethylaminonaphthalene sulfonyl chloride; OPA = o-phthalaldehyde; PITC = phenylisothiocyanate; NBD-F = 4-fluoro-7-nitrobenzo-2-oxa-l,3-diazole; HPLC = high performance liquid chromatography.

2-D Chamber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Gel Solutions and Buffers .................................................... Description of the Procedures ................................................. Microdiagonal Electrophoresis ................................................ Application................................................................ Principles of the Method .....................................................

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