LEADER 05050nam 22007094a 450 001 9910961783603321 005 20251116173937.0 010 $a9786611121150 010 $a9781281121158 010 $a1281121150 010 $a9789812708311 010 $a9812708316 035 $a(CKB)1000000000334182 035 $a(EBL)312314 035 $a(OCoLC)476099627 035 $a(SSID)ssj0000197433 035 $a(PQKBManifestationID)11172036 035 $a(PQKBTitleCode)TC0000197433 035 $a(PQKBWorkID)10154737 035 $a(PQKB)10767415 035 $a(MiAaPQ)EBC312314 035 $a(WSP)00006269 035 $a(Au-PeEL)EBL312314 035 $a(CaPaEBR)ebr10188745 035 $a(CaONFJC)MIL112115 035 $a(OCoLC)935264139 035 $a(PPN)168070251 035 $a(Perlego)849896 035 $a(BIP)13812284 035 $a(EXLCZ)991000000000334182 100 $a20071109d2007 uy 0 101 0 $aeng 135 $aur|n|---||||| 181 $ctxt$2rdacontent 182 $cc$2rdamedia 183 $acr$2rdacarrier 200 12$aA manual for biochemistry protocols /$fMarkus R. Wenk, Aaron Zefrin Fernandis 205 $a1st ed. 210 $aSingapore ;$aHackensack, NJ $cWorld Scientific$dc2007 215 $a1 online resource 225 1 $aManuals in biomedical research ;$vv. 3 300 $aIncludes index. 311 08$a9789812700667 311 08$a9812700668 320 $aIncludes bibliographical references and index. 327 $aIntro -- Contents -- Preface -- List of Figures -- List of Contributors -- A Protein Purification -- Summary -- A.1 Protein Precipitation -- A.2 Column Chromatography -- A.2.1 Ionic Exchange Chromatography -- A.2.2 Size-Exclusion Chromatography -- A.2.3 Affinity Chromatography -- A.2.4 Purification of Recombinant Proteins -- A.2.5 Commercially Prepacked Column Kits -- Notes -- B Protein Analysis -- B.1 Protein Estimation -- B.1.1 Absorbance Assays -- B.1.2 Colourimetric Assays -- B.1.3 Commercial Protein Estimation Kits -- B.2 Spectrometric Analysis -- B.3 SDS-PAGE -- B.4 Gel Staining -- B.4.1 Coomassie Blue Staining -- B.4.2 Silver Staining -- B.5 Western Blotting -- B.6 Immunoprecipitations -- Notes -- C Lipid Analysis -- C.1 Lipid Extraction -- C.1.1 Modified Bligh and Dyers Method for Phospholipid Extraction -- C.1.2 Folch Extraction -- C.1.3 Grey Method for Phosphatidylinositol Phosphate Extraction -- C.1.4 Modified Alex Brown Method for Phosphatidylinositol Phosphate Extraction -- C.1.5 Hexane Extraction for Neutral Lipids -- C.1.6 Glycolipid Extraction -- C.2 Thin Layer Chromatography -- Notes -- D Mammalian Cell Culture -- D.1 Primary Culture -- D.1.1 Primary Cell Isolation and Culturing -- D.1.2 Splenocyte Isolation -- D.1.3 Isolation of Peripheral Blood Lymphocytes -- D.2 Tissue Culture -- D.2.1 Adherent Cells -- D.2.2 Suspension Cells -- D.2.3 Cell Maintenance -- D.3 Cell Count -- D.4 Mammalian Transfection -- D.4.1 Calcium Phosphate Coprecipitation -- D.4.2 Electroporation Protocol -- D.4.3 Lipofectamine Transfection Protocol -- Notes -- E Microscopy -- E.1 Poly-L-Lysine Immobilisation -- E.2 Immunofluorescence and Confocal Microscopy -- E.3 Immunohistochemistry -- Notes -- F Assays -- F.1 Enzyme Assays -- F.1.1 Enzyme Kinetics: Basics -- F.1.2 Lipid Kinase Assay -- F.1.3 Protein Kinase Assay. 327 $aF.1.4 Protein Tyrosine Phosphatase Assay -- F.1.5 Alkaline Phosphatase Assay -- F.1.6 Caspase Assay -- F.2 Functional Assays -- F.2.1 Apoptosis Assay -- F.2.2 XTT Cell Proliferation Assay [(2,3-Bis (2-methoxy-4-nitro-5-sulfophenyl)- 2H-tetrazolium-5-carboxanilide)] -- F.2.3 Chemotaxis Assay -- F.2.4 Matrigel Invasion or Chemoinvasion Assay -- Notes -- G Autoradiography -- Summary -- Notes -- Suggested Readings -- Abbreviations -- Appendices -- Appendix 1 Safety Issues -- Appendix 2 Buffer Chart -- Appendix 3 Composition of Regular Buffers or Stock Solutions -- Index. 330 $aBiochemistry plays an important role in all areas of the biological and medical sciences. With most of the research or diagnosis involved in these areas being based on biochemically obtained observations, it is essential to have a profile of well standardized protocols. This manual is a basic guide for all students, researchers and experts in biochemistry, designed to help readers in directly starting off their experiments without prior knowledge of the protocol. The book dwells on the concepts used in designing the methodologies, thereby giving ample room for researchers to modify them according to their research requirements. 410 0$aManuals in biomedical research ;$vv. 3. 606 $aBiochemistry$vLaboratory manuals 615 0$aBiochemistry 676 $a572 676 $a572.078 700 $aWenk$b Markus R$01867463 701 $aFernandis$b Aaron Zefrin$01867464 801 0$bMiAaPQ 801 1$bMiAaPQ 801 2$bMiAaPQ 906 $aBOOK 912 $a9910961783603321 996 $aA manual for biochemistry protocols$94475015 997 $aUNINA