Protocols in Biochemistry and Clinical Biochemistry

Protocols in Biochemistry and Clinical Biochemistry, Second Edition offers clear, applied instruction in fundamental biochemistry methods and protocols, from buffer preparation to nucleic acid purification, protein, lipid, carbohydrate, and enzyme testing, and clinical testing of vitamins, glucose and cholesterol levels, among other diagnostics. Each protocol is illustrated with step-by-step instructions, labeled diagrams, and color images, as well as a thorough overview of materials and equipment, precursor techniques, safety considerations and standards, analysis and statistics, alternative methods and troubleshooting, all to support a range of study types and clinical diagnostics. This fully revised edition has been expanded and enriched to feature 100 protocols, as well as chapter key term definitions and worked examples. All-new protocols added to this edition include identification of lipids by TLC, lipid per oxidation measurement by Thiobarbituric acid (TBA) assays, Native PAGE., sub cellular fractionation of proteins, Western blotting, isoelectric focusing and 2-dimentional gel electrophoresis, protein purification, determination of serum amylase, catalase activity assay, superoxide dismutase (SOD) assay, qualitative analysis of plant secondary metabolites, qualitative analysis of photochemicals, quantitative estimation of secondary metabolites, estimation of chlorophyll contents, and starch determination, among others. Each protocol is written to help researchers and clinicians easily reproduce lab methods and ensure accurate test results. Provides updates across 100 protocols, including full listings and discussions of materials and equipment, precursor techniques, safety considerations and standards, analysis and statistics, alternative methods, and troubleshootingFeatures clear, step-by-step instruction with color diagrams and images followed by worked examples of putting lab techniques into actionEmpowers researchers and clinicians to reproduce research and clinical methods and ensure test accuracy INDICE: Unit 1: Solutions, Acids and Bases1. Preparation of Solutions2. Acid and BaseUnit 2: Carbohydrates3. Determination of the presence of carbohydrates in the given sample by Molisch’s test4. Determination of the presence of reducing carbohydrates in the given sample by Picric acid test5. Determination of the presence of reducing carbohydrates in the given sample by Fehling’s test6. Determination of the presence of reducing carbohydrates in the given sample by Benedict’s test7. Determination of the presence of reducing carbohydrates in the given sample by Tommer’s test8. Determination of the presence of reducing carbohydrates in the given sample by Nylander’s test9. Distinguish between monosaccharides and reducing disaccharides by Barfoed’s test.10. Distinguish between aldose and ketose by Seliwanoff’s test.11. Determination of pentose sugar by Bial’s test12. Determination of the presence of galactose in a test sample by the Mucic acid test13. Differentiation between the presence of ketohexose or aldohexose in the test sample by Foulger'stest14. Determination of the presence of reducing carbohydrates in the given sample by Fearon’s methylamine test15. Confirmation of the presence of reducing carbohydrates in the given sample by osazone formation test16. Determination of the presence of polysaccharides in the given sample by an iodine test17. Quantitative estimation of carbohydrates in the given sample by anthrone test.18. Quantitative estimation of reducing carbohydrates in the given sample by 3,5-dinitro salicylic acid (DNSA) test.19. Extraction and estimation of glycogen from the liver and muscle of the well-fed and starved rat.20. Separation and Identification of sugars present in fruit juices using thin-layer chromatography (TLC).21. Extraction and analysis of soluble carbohydrates from plants.Unit 3: Lipids22. Solubility test for lipids23.Emulsion test for lipids.24. To determine the degree of unsaturation of fatty acids25. Determination of the saponification value of the given fat/ oil sample26. To determine the fatty acid value of fat27. Acrolein test for the presence of glycerol28. Libermann - Buchard test for detection of cholesterol.29. Extraction of lipid from leaves (Bligh and Dyer’s method and single extraction method).30. Extraction of lipid from egg yolk and estimation of phosphorous content in it31. Extraction of lipid from tissues (Folch method).32. Identification of lipids by TLC.33. Lipid per oxidation measurement by Thiobarbituric acid (TBA) assay.Unit 4: Proteins34. Isolation of protein from animal cells/tissue using RIPA buffer35. Qualitative test for Proteins and amino acids36. Estimation of proteins by Biuret method37. Estimation of proteins by Bradford method38. Estimation of proteins by Bicinchoninic acid (BCA) method39. Estimation of proteins by Folin-Lowry method40. Estimation of proteins by UV spectrophotometric method41. Protein separation by gel electrophoresis under denaturing condition (SDS-PAGE)42. Separation of amino acids by paper chromatography.43. Native PAGE44. Sub cellular fractionation of proteins.45. Western blotting.46. Isoelectric focusing and 2-dimentional gel electrophoresis47. Protein purification.Unit 5: Minerals48. Estimation of the Calcium in water or milk by EDTA titrimetric method49. Estimation of the Calcium in serum by O-Cresolpthaleincomplexon method (colorimetric)50. Estimation of the inorganic phosphorus by colorimetric method51. Estimation of the Iron by thiocyanate colorimetric method52. Estimation of the Chloride by Titration method (Mohr Argentometric method)Unit 6: Vitamin53. Estimation of Vitamin A by Trichloroacetic acid (modified Carr-Price method)54. Estimation of the Tocopherol (Vitamin E) by colorimeter (Baker and Frank method)55. Estimation of the Thiamine (Vitamin B1) by Thiochrome technique56. Estimation of the Ascorbic acid (Vitamin C) by 2,6-dichlorophenol indophenol (DCPIP) method (Titrimetric)57. Estimation of the Ascorbic acid (Vitamin C) by 2, 4 Dinitrophenylhydrazine method (Colorimetric)Unit 7: Enzymes58. To study the effect of pH and temperature on the action of salivary amylase.59. Determination of the activity of amylase enzyme (crude) extracted from sweet potato.60. Determination of the specific activity of the amylase enzyme.61. To determine Vmax and Km in of the amylase enzyme.Unit 8: Nucleic acid62. Determination of the amount of DNA present in the solution by diphenylamine (DPA) method63. Determination of the amount of RNA present in the solution by orcinol method.64. Isolation of plasmid DNA using the alkaline lysis method.65. Preparation of plasmid DNA using boiling lysis method.66. Plasmid DNA purification using polyethylene glycol.67. Isolation of genomic DNA of the unknown bacterial strain.68. Isolation of DNA from plant tissue.69. Isolation of DNA from a eukaryotic cell using proteinase K and phenol.70. Isolation of DNA from a eukaryotic cell.71. Separation of bands of DNA by agarose gel electrophoresis.72. SOUTHERN TRANSFER AND HYBRIDIZATION: To perform Southern blotting of the genomic DNA.73. Isolation of RNA from animal cells.74. NORTHERN BLOTTING.Unit 9: Clinical Biochemistry75. Determination of Serum Urea [Diacetyl Monoxime (DAM) Method],76. Estimation of Uric Acid in Serum Sample (Uricase Method),77. Estimation of Creatinine in Serum Samples (Jaffe’s Method),78. Determination of Serum Glutamate Pyruvate Transaminase (SGPT) Activity,79. Determination of Serum Glutamate Oxaloacetate Transaminase (SGOT) Activity,80. Colorimetric Estimation of Alkaline Phosphatase (ALP) From Serum (King and Armstrong),81. Estimation of Serum Albumin by the Dye-Binding Method (Bromocresol Green Method),82. Estimation of Blood Glucose by the Glucose Oxidase Method,83. Estimation of Glucose in Urine by Benedict Method,84. Estimation of Percentage of Glucose (Reducing Sugars) in Urine (by Benedict Titration Method),85. Oral Glucose Tolerant Test (OGTT),86. Estimation of Total Cholesterol in Serum (Watson Method),87. Estimation of Total Cholesterol in Serum (Enzymatic Method),88. Estimation of Triglycerides in Serum (Enzymatic Method),89. Estimation of Hemoglobin in the Blood (Sahli’s Method).90. Determination of serum amylase.91. Catalase activity assay.92. Superoxide dismutase (SOD) assay.Unit 10: Plant Biochemistry93. Qualitative analysis of plant secondary metabolites.94. Qualitative analysis of photochemical.95. Quantitative estimation of secondary metabolites.96. Estimation of Chlorophyll contents.97. Determination of starch

• ISBN: 978-0-443-13945-1
• Editorial: Academic Press
• Encuadernacion: Rústica
• Páginas: 320
• Fecha Publicación: 01/11/2024
• Nº Volúmenes: 1
• Idioma: Inglés