LEADER 05371nam 2200637Ia 450 001 996197616503316 005 20230421044518.0 010 $a1-281-75875-2 010 $a9786611758752 010 $a3-527-61571-7 010 $a3-527-61570-9 035 $a(CKB)1000000000376034 035 $a(EBL)481439 035 $a(OCoLC)607840845 035 $a(SSID)ssj0000202309 035 $a(PQKBManifestationID)12066825 035 $a(PQKBTitleCode)TC0000202309 035 $a(PQKBWorkID)10250787 035 $a(PQKB)10199470 035 $a(MiAaPQ)EBC481439 035 $a(EXLCZ)991000000000376034 100 $a19960118d1994 uy 0 101 0 $aeng 135 $aur|n|---||||| 181 $ctxt 182 $cc 183 $acr 200 10$aMicrocharacterization of proteins$b[electronic resource] /$fR. Kellner, F. Lottspeich, H.E. Meyer 210 $aWeinheim $cVCH$dc1994 215 $a1 online resource (288 p.) 300 $aPresented at the first meeting "Mikromethoden in der Proteinchemie" held June 1994 at the Max Planck Institute for Biochemistry. 311 $a3-527-30048-1 320 $aIncludes bibliographical references. 327 $aMicrocharacterization of Proteins; Contents; Section I: Overview; 1.1 Microcharacterization of Proteins; 1 General Aspects; 2 From a Cell to a Protein Sequence; 3 FutureTrends; Section II: Sample Preparation; II.1 Chemical and Enzymatic Fragmentation of Proteins; 1 Strategy; 2 Denaturation, Reduction and Alkylation; 3 Enzymatic Fragmentation; 3.1 Enzymes; 3.2 Practical Considerations; 4 Chemical Fragmentation; 4.1 Cyanogen Bromide Cleavage; 4.2 Partial Acid Hydrolysis; 4.3 Hydroxylamine Cleavage of Asn-Gly Bonds; 4.4 Cleavage at Tryptophan; 4.5 Cleavage at Cysteine; 5 References 327 $aII.2 Microseparation Techniques I: High Performance Liquid Chromatography1 Introduction; 2 Getting Started; 2.1 Solvents; 2.2 Pump; 2.3 Pre-Column Split; 2.4 Sample Preparation; 2.5 Injector; 2.6 Tubings; 2.7 In-Line Filter, Guard Column; 2.8 Analytical Column; 2.9 Elution; 2.10 Detection; 2.11 Fractionation; 3 Applications; 4 References; II.3 Microseparation Techniques II: Analysis of Peptides and Proteins by Capillary Electrophoresis; 1 Introduction; 2 Theory; 2.1 Capillary Isotachophoresis; 2.2 Capillary Zone Electrophoresis; 2.3 Electroosmotic Flow; 3 Instrumentation; 3.1 Injection 327 $a3.2 Detection4 Applications; 4.1 Peptide Separations; 4.2 Protein Separations; 5 References; II.4 Microseparation Techniques III: Gel Electrophoresis for Sample Preparation in Protein Chemistry; 1 Introduction; 2 Denaturing Techniques; 2.1Commonly Used SDS-Polyacrylamide Gel Electrophoresis Techniques for Protein Separation; 2.2 Blue-SDS-PAGE for Quantitative Protein Recovery from Gels; 2.3 Electroelution of Proteins After Blue-SDS-PAGE; 2.4 Electroblotting of Blue and Colourless SDS Gels; 2.5 Isoelectric Focusing in the Presence of Urea; 3 Native Techniques; 3.1 Colourless-Native-PAGE 327 $a3.2 Blue-Native-PAGE3.3 Native Isoelectric Focusing; 4 References; II.5 Microseparation Techniques IV: Electroblotting; 1 Introduction; 2 Electroblotting; 2.1 Polyacrylamide Gel Electrophoresis; 2.2 Blot Systems; 2.2.1 Tank Blotting; 2.2.2 Semidry Blotting; 2.3 Blotting Parameters; 2.3.1 The Blotting Process; 2.3.2 Transfer Buffers; 2.3.3 Addition of SDS; 2.3.4 Addition of Methanol; 2.3.5 Influence of Protein Concentration; 3 Blotting Membranes; 4 References; Section III: Amino Acid Analysis; III.1 Amino Acid Analysis; 1 Introduction; 2 Sample Preparation; 2.1 Peptides and Proteins 327 $a2.1.1 Enzymatic Hydrolysis2.1.2 Acid Hydrolysis; 2.1.3 Alkaline Hydrolysis; 2.2 Free Amino Acids; 3 Derivatization; 3.1 Post-Column Derivatization; 3.1.1 Ninhydrin; 3.1.2 Orthophthaldialdehyde; 3.1.3 Fluorescamine; 3.2 Pre-Column Derivatization; 3.2.1 Phenylisothiocyanate; 3.2.2 Orthophthaldialdehyde; 3.2.3 Fluorenylmethyl Chloroformate; 3.2.4 Dabsyl Chloride; 3.2.5 Dansyl Chloride; 3.2.6 Chiral Reagents; 4 Data Evaluation; 5 Instrumentation; 6 Discussion; 7 References; Section IV. Protein Sequence Analysis; IV. 1 The Edman Degradation; 1 The Edman Chemistry 327 $a1.1 Coupling, Cleavage and Conversion 330 $aBoth a thorough introduction and laboratory guide!This book is based on the lectures and demonstrations presented at a successful workshop organized by the authors. Expert contributions and the support of more than a dozen companies engaged in bioanalysis and instrumentation enabled the participants to familiarize themselves with the most recent developments e.g. in protein separation and characterization (including laser desorption-ionisation mass spectrometry), fragmentation and micro sequencing.The workshop was held in the Max Planck Institute for Biochemistry at Martinsried 606 $aProteins$vCongresses 606 $aProteins$xAnalysis$vCongresses 615 0$aProteins 615 0$aProteins$xAnalysis 676 $a547.75046 676 $a572/.65 700 $aKellner$b R$g(Roland)$0737934 701 $aLottspeich$b F$g(Friedrich),$f1947-$01231893 701 $aMeyer$b H. E$g(Helmut E.)$01231894 801 0$bMiAaPQ 801 1$bMiAaPQ 801 2$bMiAaPQ 906 $aBOOK 912 $a996197616503316 996 $aMicrocharacterization of proteins$92860413 997 $aUNISA