Practical Biochemistry

Name: Practical Biochemistry
Brand: De Gruyter
Price: 53.6 EUR
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Pamela Jha(Autor*in)

De Gruyter (Verlag)

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Erschienen am 26. August 2024

330 Seiten

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978-981-5165-85-2 (ISBN)

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Inhalt

Cover
Title
Copyright
End User License Agreement
Contents
Foreword
Preface
List of Contributors
Introduction to Carbohydrates
Mrittika Sarkar1, Sinchan Hait2,* and Sai Joshi3
INTRODUCTION
Classification of Carbohydrates
Monosaccharides (mono = one)
Oligosaccharides (oligo = few)
Polysaccharides (poly = many)
GLYCOSIDIC BOND
N-glycosides
C-glycosides
S-glycosides
O-glycosides
REDUCING AND NON-REDUCING SUGARS
PROPERTIES OF CARBOHYDRATES
Elemental Composition
Physical Properties
Stereoisomers
Geometrical (cis-trans isomers)
Enantiomers
Optical Activity
Epimers
Anomers
CONCLUSION
VIVA VOCE
REFERENCES
Qualitative Tests for Carbohydrate Detection
Mrittika Sarkar1, Sinchan Hait2,* and Sai Joshi3
INTRODUCTION
Solubility Test
Aim
Principle
Requirements
Procedure
Applications
Conclusion
Molisch's Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Applications
Fehling Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Applications
Benedict's Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Applications
Seliwanoff's Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Troubleshooting
Applications
Bial's Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Application
Iodine Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Troubleshooting
Application
Barfoed's Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Application
Osazone Test
Aim
Principle
Requirements
Procedure
Conclusion
Precautions
Troubleshooting
Application
DID YOU KNOW?
CONCLUSION
VIVA VOCE
REFERENCES
Quantification Tests for Carbohydrate Detection
Mrittika Sarkar1, Anshika Sah2, Payal Agarwal3, Sai Joshi4 and Sinchan Hait5,*
INTRODUCTION
ANTHRONE METHOD
Principle
Hydrolysis of Complex Carbohydrates
Dehydration of Monosaccharide
Reaction with Reagent
Requirements
Procedure
Results
Did you know?
DNSA METHOD
Principle
Requirements
Procedure
Results
Did you know?
FOLIN-WU METHOD
Principle
Requirements
Procedure
Results
HEXOKINASE METHOD
Principle
Requirements
Procedure
Results
PHENOL-SULFURIC ACID METHOD
Principle
Requirements
Procedure
Results
SOMOGVI-NELSON METHOD
Principle
Requirements
Procedure
Results
GOPOD METHOD
Principle
Requirements
Procedure
Results
CONCLUSION
VIVA VOCE
REFERENCES
Introduction to Proteins
Sinchan Hait1,* and Sai Joshi2
INTRODUCTION
PROTEIN STRUCTURE
FUNCTIONS OF PROTEIN
Enzymes
Membranes and Transport Molecules
Antibodies and Toxins
Functional Proteins
Structural Proteins
CONCLUSION
VIVA VOCE
REFERENCES
Qualitative Tests for Protein Detection
Sinchan Hait1,*
INTRODUCTION
AROMATIC AMINO ACIDS (TYROSINE, TRYPTOPHAN)
Xanthoproteic Test
Aim
Principle
Requirement
Procedure
Results
Precaution
Application
Limitations
TYROSINE
Millon's Test
Aim
Principle
Requirement
Procedure
Results
Precaution
Application
Limitations
TYROSINE AND HISTIDINE
Pauly's Diazo Test
Aim
Principle
Requirement
Procedure
Results
Precaution
Application
Limitation
TRYPTOPHAN
Hopkin's-Cole Test
Aim
Principle
Requirement
Procedure
Results
Precautions
Application
Limitation
Ehrlich's Test
Aim
Principle
Requirement
Procedure
Results
Applications
Limitations
Did you know?
ARGININE
Sakaguchi Test
Aim
Principle
Requirement
Procedure
Results
Application
Limitation
CYSTEINE
Lead Sulfide Test
Aim
Principle
Requirement
Procedure
Results
Application
Limitation
Nitroprusside Test
Aim
Principle
Requirement
Procedure
Results
Precautions
Applications
Limitations
Ninhydrin Test
Aim
Principle
Requirement
Procedure
Results
Precaution
Application
Limitation
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Protein Determination by SDS-PAGE
Sai Joshi1,*
INTRODUCTION
PRINCIPLE
REAGENT PREPARATION
REQUIREMENTS AND PROCEDURES
Preparation of Gel
Requirements
Procedure
Preparation of Samples
Requirements
Procedure
Loading the Sample
Requirements
Procedure
Running the Gel
Requirements
Procedure
Staining the Gel
Part A - Coomassie Blue Staining
Part B - Silver Staining
Destaining the Gel
Requirements
Procedure
Visualization of the Gel
PRECAUTION
APPLICATIONS
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Protein Determination by Western Blotting or Immunoblot
Sinchan Hait1,*
INTRODUCTION
QUALITATIVE WESTERN BLOTTING
Principle
Did you know?
Requirement
Procedure
SDS-PAGE
Electrotransfer
Blocking and Antibody Incubation
Detection
QUANTITATIVE WESTERN BLOTTING
Principle
Interpretation of Test Results
Positive Result
Negative Result
Invalid
Precautions
Troubleshooting
Unexpected Bands
Blurred Bands
No Bands
High Background
Uneven Bands on Blot
CONCLUSION
VIVA VOCE
REFERENCES
Quantitative Tests for Protein Detection
Sinchan Hait1, Sai Joshi2,* and Ankita Nitin Padhye3
INTRODUCTION
BICINCHONINIC ACID (BCA) TEST
Aim
Principle
Requirements
Procedure
Standard Bicinchoninic Acid Preparation
Standard Bicinchoninic Acid Preparation (For micro-assay)
MacroAssay
MicroAssay (0.5-10 pg/ml)
Results
Results Interpretation
Troubleshooting
Did you know?
BIURET METHOD
Aim
Principle
Requirement
Procedure
Preparation of Biuret Reagent
Results
Advantages
Disadvantages
Troubleshooting
Did you know?
BRADFORD ASSAY
Aim
Principle
Requirements
Reagents
Protein Solutions
Procedure
Protein Preparation
Preparation of Bradford Reagent
Protein Assay (Standard Method)
MicroProtein Assay
Results
Applications
Troubleshooting
Advantages
Disadvantages
Did you know?
LOWRY METHOD
Aim
Principle
Requirements
Reagent I: Lowry Reagent (or) Alkaline Copper Sulfate Solution
Reagent II: Folin-Ciocalteau Reagent
Standard Protein Solution
Procedure
Results
Troubleshooting
Applications
Did you know?
ADVANCED TECHNIQUES FOR PROTEIN QUANTIFICATION
Fluoroprofile
NanoOrange Fluorescence Protein Estimation
Accu Orange Fluorescence Protein Estimation
CONCLUSION
VIVA VOCE
REFERENCES
Protein Purification
Sinchan Hait1, Sai Joshi2,* and Ankita Nitin Padhye3
INTRODUCTION
PURIFICATION OF DIFFERENT TYPES OF PROTEINS (WITHOUT TAG)
Ammonium Sulfate Precipitation
Aim
Principle
Requirements
Procedure
Results
Precautions
Advantages
Limitations
Protein Purification by Dialysis
Aim
Principle
Factors Affecting Dialysis
Temperature
Concentration and Molecular Weight of the Molecule
Surface Area and Thickness of the Membrane
Requirements
Procedure
Dialysis Process
Precautions
Troubleshooting
Did you know?
PURIFICATION OF DIFFERENT TYPES OF RECOMBINANT PROTEINS (WITH TAG)
Protein Purification by Affinity Chromatography
Aim
Principle
Components in Affinity Chromatography
Affinity Tag Removal
Functions of Protein Tags
Reasons for Tag Removal
Basic Steps Involved in the Removal of the Tag
Requirements
Procedure
Precautions
Advantages
Disadvantages
Protein Purification by Size Exclusion Chromatography
Aim
Principle
Application
Types of Gel Filtration Chromatography
Apparatus
Procedure
Precautions
Advantages
Disadvantages
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Introduction to Nucleic Acids
Sai Joshi1,*, Sinchan Hait2 and Ahiree Ghosal3
INTRODUCTION
Deoxyribonucleic Acid (DNA)
Functions of DNA
Components of DNA
RNA: Ribonucleic Acid
Ribosomal RNA (rRNA)
Messenger RNA (mRNA)
Transfer RNA (tRNA)
Components of RNA
The Structure of Nucleic Acids
The Establishment of DNA as Genetic Information
Isolation of DNA
Evidence that DNA is the Genetic Material
DNA Accepted as the Genetic Material
Erwin Chargaff Rules
DNA X-ray Fiffraction Pattern
Qualitative Analysis of Nucleic Acids
Quantitative Analysis of Nucleic Acids
Spectrophotometric Analysis
Fluorometric Methods for Nucleic Acid Quantitation
Specificity
Sensitivity
Dynamic Range
Sample Volume
Sample Format
Instrumentation Filters
Capillary Electrophoresis
CONCLUSION
VIVA VOCE
REFERENCES
Qualitative Tests for Nucleic Acid Detection
Anshika Sah1, Mrittika Sarkar2, Sinchan Hait3, Sai Joshi4,* and Ahiree Ghosal5
INTRODUCTION
AGAROSE GEL ELECTROPHORESIS
Aim
Principle
Reagents and Materials
DNA Electrophoresis
RNA Electrophoresis
Procedure
DNA
RNA
Precautions
Gel A
Gel B
Troubleshooting
Gel C
Troubleshooting
Gel D
Troubleshooting
NUCLEIC ACID QUALITY AND QUANTITY ESTIMATION BY NANODROP SPECTROMETRY
Principle
Requirements
Procedure
Results
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Quantitative Tests for Nucleic Acid Detection
Mrittika Sarkar1, Sinchan Hait2, Sai Joshi3,* and Revati Supekar4
INTRODUCTION
SPECTROPHOTOMETRY OF DNA AND RNA
Aim
Principle
Procedure
Troubleshooting
COLOURIMETRIC ASSAYS
Diphenylamine Reaction
Aim
Principle
Procedure
Orcinol Reaction
Aim
Principle
Procedure
Precautions
QUANTIFICATION OF NUCLEIC ACIDS BY ADVANCED FLUORESCENCE TECHNIQUES
DNA Detection using the DNA-binding Fluorochrome Hoechst 33258:
Principle
Advantages
Disadvantages
DNA And RNA Detection with Ethidium Bromide Fluorescence
Principle
Advantages
Disadvantages
DNA Detection using DNA-binding Fluorochrome PicoGreen
Principle
Requirements
Procedure
Results
Q-PCR
Principle
Requirements
Procedure
Results
Advantage
Disadvantages
CONCLUSION
VIVA VOCE
REFERENCES
Purification of Nucleic Acids
Sai Joshi1,*, Sinchan Hait2 and Niriksha Patel3
INTRODUCTION
SEPARATION OF SOLUBLE DNA FROM CELL DEBRIS AND OTHER INSOLUBLE MATERIAL
Extraction of the Nucleic Acid Sample by Phenol-chloroform Method
Principle
Binding to the Purification Matrix
Solution-Based Chemistry
ALCOHOL-BASED PURIFICATION METHOD
Aim
Principle
Requirements
Procedure
Precaution
Did you know?
Silica-Binding Chemistry
SILICA COLUMN-BASED PURIFICATION
Aim
Principle
Requirements
Glasswares
Other Requirements
Reagents
Preparation of Reagents
Procedure
Preparation of DNA Solution
Purifying DNA on Silica Membrane Spin Column
Results
Precautions
Did you know?
Cellulose-Binding Chemistry
AGAROSE GEL ELECTROPHORESIS BASED PURIFICATION
Aim
Principle
Requirements
Glasswares
Other Requirements
Reagents
Preparation of Reagents
Procedure
Preparation of Gel
Setting up of Gel Apparatus and Separation of DNA Fragments
Visualizing Separated DNA Fragments
Results
Precautions
Did you know?
Ion Exchange Chemistry
CONCLUSION
VIVA VOCE
REFERENCES
Introduction to Lipids
Sai Joshi1, Sinchan Hait2,* and Pranali Garde3
INTRODUCTION
Fats
Glycerol
Fatty acids
Structure of Fatty Acids
Classification on the Basis of Saturation
Saturated Fatty Acids
Unsaturated Fatty Acids
Fatty Acid Derivatives
Triglycerides
Waxes
Glycerophospholipids
Sphingolipids
Cholesterol and its Derivatives
Lipoproteins
Classification and Formation
Chylomicrons
Very Low-Density Lipoproteins (VLDL)
Low-Density Lipoprotein (LDL)
High-Density Lipoprotein (HDL)
Eicosanoids
Advanced Techniques used for Lipid Quantification
Conventional Method
Non-conventional Method
Criteria for Selection of Advanced Method for Lipid Quantification
Gravimetric Analysis
Chromatographic Analysis
Spectroscopic Analysis
Colourimetric Analysis
CONCLUSION
VIVA VOCE
REFERENCES
Qualitative Tests for Lipid Detection
Sinchan Hait1,*, Sai Joshi2 and Pranali Bhosale3
INTRODUCTION
SOLUBILITY TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
TRANSLUCENT SPOT TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
EMULSIFICATION TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
SAPONIFICATION TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
SUDAN IV TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
ACROLEIN TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
DICHROMATE TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
TEST FOR FREE FATTY ACIDS
Aim
Principle
Requirements
Procedure
Observation
Inference
Troubleshooting
UNSATURATION TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
BURCHARD TEST
Aim
Principle
Requirements
Procedure
Observation
Inference
Precautions
DID YOU KNOW?
CONCLUSION
VIVA VOCE
REFERENCES
Quantitative Tests for Lipid Detection
Sai Joshi1, Sinchan Hait2,* and Pranali Garde3
INTRODUCTION
GRAVIMETRIC ANALYSIS
Principle
Requirements
Procedure
Observation
Determination of Total Lipid Content
Results
Precaution
Application
Advantages
Disadvantages
ACID VALUE
Aim
Principle
Requirements
Procedure
Calculation
Result
Precaution
Application
Advantages
Disadvantages
SAPONIFICATION VALUE
Aim
Principle
Requirements
Procedure
Calculation
Result
Precaution
Application
Advantages
Disadvantages
Did You Know?
IODINE VALUE
Aim
Principle
Requirements
Procedure
Calculation
Results
Precaution
Application
Advantages
Disadvantages
Did You Know?
BLOOD CHOLESTEROL
Aim
Principle
Requirements
Procedure
Calculation
Results
Precaution
Advantages
Disadvantages
Did You Know?
ADVANCED TECHNIQUES OF LIPID QUANTIFICATION
Chromatographic Analysis
Thin Layer Chromatography
Did you know?
Gas Chromatography
Did you know?
High-performance Liquid Chromatography of Lipids
Spectroscopic Analysis
Infrared Spectroscopy
Did you know?
Nuclear Magnetic Resonance Spectroscopy
Did you know?
Raman Spectroscopy
Did you know?
Fluorescence Spectroscopy
Did you know?
Colourimetric Analysis
Aim
Principle
Requirements
Storage and Stability
Procedure
Calculation
Normal Value
Results
Precautions
Troubleshooting
Application
Advantages
Disadvantages
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Commonly Used Basic and Advanced Techniques
Anshika Sah1, Sinchan Hait2 and Sai Joshi3,*
INTRODUCTION
MEASUREMENT OF REDOX POTENTIAL
Aim
Principle
Requirement
Procedure
Results
Precaution
Advantages
Disadvantages
Troubleshooting
MEASUREMENT OF PH
Principle
Requirements
Procedure
Calibration
pH Measurement
Results
Advantages
Disadvantages
Troubleshooting
Precautions
MEASUREMENT OF ZETA POTENTIAL
Principle
Measurement of Zeta Potential
Requirements
Procedure
Advantages
Disadvantages
Troubleshooting
Precautions
SEPARATION USING HPLC
Principle
Requirements
Instruments
Procedure
Sample Reparation
HPLC Method Development
Results
Troubleshooting
Precautions
Advantages
Disadvantages
ION-EXCHANGE CHROMATOGRAPHY
Anion Exchange
Principle
Purification of Nucleic Acid
Requirements
Pump
Injector
Column
Suppressor
Detector
Data System
Procedure
Applications
Did you know?
CONCLUSION
VIVA VOCE
REFERENCES
Significant Experimental Hazard: Safety Data Sheet (SDS)
Sai Joshi1,*, Anshika Sah2 and Sinchan Hait3
INTRODUCTION
Section 1
Section 2
Section 3
Section 4
Section 5
Section 6
Section 7
Section 8
Section 9
Section 10
Section 11
Section 12
Section 13
Section 14
Section 15
Section 16
HAZARD INFORMATION ON CHEMICALS USED IN THE EXPERIMENTS FROM SDS
Eye Damage
Respiratory Difficulties
Skin Damage
Ingestion
CONCLUSION
VIVA VOCE
REFERENCES
Subject Index
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