Tuesday, July 6, 2010

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    Sunday, March 7, 2010

    The ELISA Guidebook

    The ELISA Guidebook
    By John R. Crowther

    Publisher: Humana Press

    Number Of Pages: 436

    Publication Date: 2000-08

    Sales Rank: 282361

    ISBN / ASIN: 0896037282

    EAN: 9780896037281

    Binding: Paperback

    Manufacturer: Humana Press

    Book Description:


    John R. Crowther provides todayís premier practical guide to the understanding and application of ELISA. Updating and greatly expanding his widely appreciated earlier publication, ELISA Theory and Practice (1995), this important work introduces chapters on such major new topics as checkerboard titrations, quality control of testing, kit production and control, novel monoclonal antibodies, validation of assays, statistical requirements for data examination, and epidemiological considerations. With its numerous worked examples, detailed instructions, and extensive illustrations, The ELISA Guidebook offers a powerful synthesis of all the basic concepts and practical experimental details investigators need to understand, develop, and apply the new ELISA methodology successfully in day-to-day basic and clinical research

    http://rapidshare.com/files/140990550/the_elisa_guide_book-spiderman.pdf


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    Monday, February 22, 2010

    Harper's Biochemistry 28th edition

    Harper's Illustrated Biochemistry, 28th Edition : McGraw-Hill Professional Online Book Store

    Overview

    The biochemistry text that every medical student must own--now in full color!

    Comprehensive, concise, and up-to-date, Harper's is unrivaled in its ability to clarify the link between biochemistry and the molecular basis of health and disease.

    The Twenty-Eighth Edition has undergone sweeping changes -- including a conversion to full-color artwork and the substantial revision and updating of every chapter -- all to reflect the latest advances in knowledge and technology and to make the text as up-to-date and clinically relevant as possible. Combining outstanding full-color illustrations with integrated coverage of biochemical diseases and clinical information, Harper's Illustrated Biochemistry offers an organization and clarity not found in any other text on the subject.

    Striking just the right balance between detail and brevity, Harpers Illustrated Biochemistry is essential for USMLE review and is the single best reference for learning the clinical relevance of a biochemistry topic.

    NEW to this edition:

    * Full-color presentation, including 600+ illustrations
    * Every chapter opens with a Summary of the Biomedical Importance and concludes with a Summary reviewing the topics covered
    * Two all-new chapters: "Free Radicals and Antioxidant Nutrients" and "Biochemical Case Histories" which offers an extensive presentation of 16 clinical conditions
    * A new appendix containing basic clinical laboratory results and an updated one with a list of important websites and online journals
    * NEW or updated coverage of important topics including the Human Genome Project and computer-aided drug delivery

    Table of contents

    1. Biochemistry and Medicine
    2. Water & PH
    3. Amino Acids and Peptides
    4. Proteins: Determination of Primary Structure
    5. Proteins: Higher Orders of Structure
    6. Proteins: Myoglobin and Hemoglobin
    7. Enzymes: Mechanisms of Action
    8. Enzymes: Kinetics
    9. Enzymes: Regulation of Activities
    10. Bioenergetics: The Role of ATP
    11. Biologic Oxidation
    12. The Respiratory Chain & Oxidative Phosphorylation
    13. Carbohydrates of Physiologic Significance
    14. lipids of Physiologic Significance
    15. Overview of Metabolism
    16. The Citric acid Cycle: The Catabolism of Acetyl-CoA
    17. Glycolysis and Oxidation of Pyruvate
    18. Metabolism of Glycogen
    19. Gluconeogenesis and Control of the Blood Glucose
    20. The Pentose Phosphate Pathway & other Pathways of Hexose Metabolism
    21. Biosynthesis of Fatty Acids
    22. Oxidation of Fatty Acids: Ketogenesis
    23. Metabolism of Unsaturated Fatty Acids & Eicosanoids
    24. Metabolism of Acyglycerols and Sphingolipids
    25. Lipid Transport And Storage
    26. Cholesterol Synthesis, Transport and Excretion
    27. Integration of Metabolism- the Provision of Metabolic Fuels
    28. Biosynthesis of Nutritionally Nonessential Amino Acids
    29. Catabolism of Proteins & of Amino Acid Nitrogen
    30. Catabolism of the Carbon Skeletons of Amino Acids
    31. Conversion of Amino Acids
    32. Porphyrins & Bile Pigments
    33. Nucleotides
    34. Metabolism of Purine & Pyrimidine Nucleotides
    35. Nucleic Acid Structure and Function
    36. DNA Organization, Replication and Repair
    37. RNA Synthesis, Processing and Modifications
    38. Protein Synthesis and Genetic Code
    39. Regulation of Gene Expression
    40. Molecular Genetics, Recombinant DNE and Genomic Technology
    41. Membranes: Structure and Function
    42. The Diversity of Endocrine System
    43. Hormone Action and Signal Transduction
    44. Nutrition, Digestion and Absorption
    45. Vitamins and Minerals
    46. Intracellular Traffic and Sorting of Proteins
    47. Glycoprotein
    48. The Extracellular Matrix
    49. Muscle and the Cytoskeleton
    50. Plasma Proteins and Immunoglobulins
    51. Homeostasis and Thrombosis
    52. The Red and White Blood Cells
    53. Metabolism of Xenobiotics
    54. The Human Genome Project
    Biographical note

    Robert K. Murray, MD, PhD,
    Professor (Emeritus) of Biochemistry,
    University of Toronto

    David A. Bender, PhD,
    Sub-Dean, University College Medical School,
    Senior Lecturer in Biochemistry,
    Department of Structural and Molecular Biology and
    Division of Medical Education,University College London

    Kathleen M. Botham, PhD, DSc,
    Professor of Biochemistry, Royal Veterinary
    College, University of London

    Peter J. Kennelly, PhD,
    Professor and Head, Department of Biochemistry,
    Virginia Polytechnic Institute and State University,
    Blacksburg, Virginia

    Victor W. Rodwell, PhD,
    Emeritus Professor of Biochemistry, Purdue University, West Lafayette,
    Indiana

    P. Anthony Weil, PhD,
    Professor of Molecular Physiology and Biophysics,
    Vanderbilt University School of Medicine,
    Nashville, Tennessee

    Co-Authors:

    Daryl K. Granner, MD,
    Emeritus Professor of Molecular Physiology and Biophysics
    and Medicine,
    Vanderbilt University, Nashville, Tennessee
    Peter L. Gross MD, MSc, FRCP(C),
    Associate Professor, Department of Medicine,
    McMaster University

    Frederick W. Keeley, PhD,
    Associate Director and Senior Scientist, Research Institute,
    Hospital for Sick Children, Toronto, and Professor,
    Department of Biochemistry, University of Toronto

    Peter A. Mayes, PhD, DSc,
    Emeritus Professor of Veterinary Biochemistry, Royal Veterinary College, University of London

    Margaret L. Rand, PhD,
    Associate Senior Scientist, Hospital for Sick Children, Toronto,
    and Professor, Departments of Laboratory Medicine and Pathobiology
    and Department of Biochemistry, University of Toronto

    download link
    http://rapidshare.com/files/282803148/H_BC_28ed_lycan.rar

    torrent link
    http://freshwap.net/torrents/dl/2991880





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    Monday, February 15, 2010

    Carbohydrate Chemistry

    Introduction 

             Carbohydrates have the general molecular formula (CH2O)n, and thus were once thought to represent "hydrated carbon".

             All sugars are very soluble in water because of their many hydroxyl groups.

             They are called sugars because they are sweet in taste & crystalline in nature.

             Sugars are the most important source of energy for many cells.

    Definition:

             Hydrates of carbon Cn(H2O)n

    o       {exceptions:

             Acetic acid, C2H4O2 & lactic Acid, C3H6O3 Formaldehyde, CH2Oare not carbohydrates but

             Deoxy Ribose C6H12O5 is a carbohydrate

             carbohydrates are polyhydroxy aldehydes or ketones or compounds, which yields these on hydrolysis.

    Classification:

             Hyaluronic acid

             Heparin

             Chondroitin SO4

             Keratan SO4
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    Carbohydrates

    CARBOHYDRATES

    Carbohydrates have the general molecular formula (CH2O)n, and thus were once thought to represent "hydrated carbon". All sugars are very soluble in water because of their many hydroxyl groups. They are called sugars because they are sweet in taste & crystalline in nature. Although not as concentrated a fuel as fats, sugars are the most important source of energy for many cells. Carbohydrates provide the bulk of the calories (4 kcal/gram)

    1)Definition:

    a)Hydrates of carbon Cn(H2O)n{exceptions: Acetic acid, C2H4O2 & lactic Acid, C3H6O3 Formaldehyde, CH2Oare not carbohydrates but Rahnose,C6H12O5 is.}

    b)carbohydrates are polyhydroxy aldehydes or ketones or compounds, which yields these on hydrolysis.

    2)Classification:

    a)Monosaccharides:simple sugars that cannot be hydrolyzed to yield Carbohydrates

    Name

    Aldose

    Ketose

    Formula

    Trioses

    Glyceraldehyde

    Dihydroxy acetone

    C3H6O3

    Tetroses

    Erythrose

    Erytrulose

    C4H8O4

    Pentoses

    Ribose

    Ribulose

    C5H10O5

    Hexoses

    Glucose,Mannose, Galactose

    Fructose

    C6H12O6

    Heptoses

    Sedoheptulose

    C7H14O7

    Nonoses

    Sialic acid or Neuramic acid

    C9H18O9

    b)Disaccharides: yields two molecules of Monosaccharides on hydrolysis

    Name

    Monosaccharides

    Formula

    Sources

    Properties

    Maltose

    Glucose + Glucose

    C3H6O3

    Malt sugar, starch

    Reducing

    Lactose

    Glucose + Galactose

    C3H6O3

    Milk Sugar

    Reducing

    Sucrose

    Glucose + Fructose

    C3H6O3

    Cane sugar

    Non-Reducing

    Sucrose: prevalent in sugar cane and sugar beets, is composed of glucose and fructose through an a-(1,2)b-glycosidic bond.

    Lactose: is found exclusively in the milk of mammals and consists of galactose and glucose in a b-(1,4) glycosidic bond.

    Maltose: the major degradation product of starch, is composed of 2 glucose monomers in an a-(1,4) glycosidic bond.

    c)Oligosaccharides: yields 3 to 10 Monosaccharides on Hydrolysis.

    Maltotriose

    Glucose + Glucose + Glucose

    Raffinose

    A trisaccharide that occurs in sugar beets and cotton seeds and cereals

    Stachyose

    A tetrasaccharide found in the tubers of the Chinese artichoke

    d)Polisaccharides: yields more than 10 Monosaccharides on hydrolysis. They are of two types.

    i)Homopolysaccharides (Homoglycans) Polymer of a single type of Monosaccharide.

    Name

    Monosacc.

    Sources

    Importance

    Starch

    Glucose

    Potato,tapioca,rice,wheat,&grains

    Plant source of Energy

    Amylose

    Glucose

    Un branched starch similar to cellulose, containing alpha(1->4) linkages

    Amylopectin

    Glucose

    Branched starch containing both 1,4 and 1,6 linkages.

    Glycogen

    Glucose

    5% of liver mass> branched & compact

    Animal storage of Glucose

    Inulin

    Fructose

    various bulbs&tubers(onion,garlic)

    Renal clearance studies.

    Dextrins

    Glucose

    Product. ofstarch digestion

    White dextrin is used in pharmaceutical preparations

    Chitin

    N-Ac-Glucosamine

    Exoskeleton of Insects


     

    ii)Hetroplysaccharides (Hetroglycans): polymer of two or more types of sugar residues

    Agar

    Glu +Gal + other

    Obtained from Sea weeds

    A colloidal extract of algae; used especially in culture media and as a gelling agent in foods (undigested by bacteria, dissolves at 100 oC and upon cooling sets into a Gel)

    iii)MUCOPOLYSACCHARIDES (MPS) or GLYCOSAMINOGLYCANS (GAG)

    Are Carbohydrates containing Uronic Acid & Amino Sugars.

    Name

    Functions

    Hyaluronic acid

    A viscous mucopolysaccharide found in the connective tissue space and the synovial fluid of movable joints and the humours of the eye; a cementing and protective substance

    Heparin

    A polysaccharide produced in basophils (especially in the lung and liver) and that inhibit the activity of thrombin in coagulation of the blood.

    Chondroitin SO4

    Inground substance of connective tissues (cartilage, tendon, bone)

    Keratan SO4

    In cornea & tendons.


     

    e)Isomerism

    i)D & L Isomerism

    oThe configuration of the asymmetric carbon atom ( carbon atom whose 4 valencies are filled up by 4 different groups)farthest from the aldehydic or ketonic group

    oIf –OH group is attached to the right then 'D' series & left then 'L' series.

    ii)Pyranose - Furanose ring Structure

    oWhen ring structure were suggested by Haworth (projections)

    oFive member ring structure is calledFuranose and Six number as Pyranose

    iii)Alpha & Beta Anomerism

    oThe anomeric carbon (which originally was a part of the aldo or keto group) forms a hemiacetal or hemiketal in the ring structure generating a new asymmetric centre.

    oIf hydroxyl gp (-OH) of the anomeric carbon is to the right then it is -form and if to the left it -form.

    iv)Epimerism

    oEpimers differ in the position of the Hydroxyl gp at only one of their assymetric C-atom.

    oD-Glucose & D-Galactsoe are C-4 Epimers

    oD-Glucose & D-Mannose are C-2 epimers

    v)Enantiomers

    oStereo isomers that are mirror images of each other

    oEg: D-Glyceraldehyde & L-Glyceraldehyde.

    vi)Mutarotation

    oWhen D-Glucose is crystallized at RT and fresh solution is prepared, its specific rotation of polarized light is +112 o; but after few hrs it changes to +52.5 o. This change in rotation is called Mutarotation. This b'se D-Glucose has two anomers & forms. (-D-Glucose has specific rotation of +112 o and -D-Glucose has +19 o at equilibrium 1/3 is form and 2/3 is form to get a specific rotation of +52.5 o

    3)Other Physiologically important carbohydrates.

    Sugar

    Present where

    Importance&clinical Significance

    D-Ribose

    Nucleic acids

    Nucleic acid co-enzymes.

    D-Ribulose

    Metabolic Product

    Pentose-PO4 pathway intermediate.

    D-Arabinose

    Gum arabic, Plum & Cherry gum

    Constituent of Glycoproteins

    D-Xylose

    Wood gums, GAG's

    Constituent of Glycoproteins

    D-Xylulose

    Intermediate in Uronic acid pathway

    Found in the Urine of patients with Essential pentosuria


    4)CHEMISTRY

    a)Reactions

    i)Benedict's Reaction

    Contains Na2CO3, Na-Citrte,CuSO4

    In alkaline medium (Na2CO3), the copper remains as cupric hydroxide in presence of sodium citrate

    5 ml Benedict's reagent+ 0.5 ml Urine boil for 2 minutes. If sugar present copper is reduced to produce green, yellow, orange, red precipitate depending on the concentration of the sugar.

    SugarEnediolCu++(cupric)  CuSO 4(Reagent)

    Sugar AcidCu+(cuprous) 2Cu(OH)[Yellow]Cu 2O [Red]

    Chemical propertie

    s

    1

    Reactions of aldehyde / keto group

      

    Reactions

    Reagents

    Result

     

    a.

    Osazone formation

    Sugar+Phenyl Hydrazine HCl+ Sodium Acetate (boil+cool)

    Glucosazone-needle shapedLactosazone - Powder puff Maltosazone - Sunflower

    b.

    Reduction → Alcohol

    Sodium amalgum / Hydrogen

    Glucose - sorbitolFructose - Manitol

    c.

    Oxidation

    		  

    I.

    Mild (CHO → COOH)

    Br2 H2O

    Glucose - gluconic acid

    ii.

    Srong (CHO & CH2OH to COOH

    HNO3

    glucose - Glucosaccharic acid

    iii.

    Oxdn of CH2OH alone

    Enzymatic (living cells only)

    Glucose to glucuronic acid

    d.

    Tautomerisation( Inter conversions)

    Weak alkaliBa (OH)2, Ca(OH)2

    Glucose ↔ fructose ↔ Mannose

    e.

    Reduction Reactions

    CuSO4, alkaline medium( cupric -> cuprous)

    Benedicts, Fehlings & Barfoeds Test( Cuprous oxide- Red ppt)

    2

    Reactions of Alcoholic group

     

    a.

    Glycosides

    1→4 &1→6 bond in glycosidic bonds in glycogen

      

    I.

    Cardiac

    Derivatives of DIGITALIS,

    Drugs in cardiac insufficiency

     

    ii.

    Oubin

    strophanthus sp.

    Sodium pump inhibitor (cardiac)

     

    iii.

    Phlorbizin

    Root/bark of apple tree

    block transport of sugar

     

    iv.

    Streptomycine

    produced by the actinomycete Streptomyces griseus

    Antibioticused to treat tuberculosis

     

    b.

    Acetilation/Ester formation

    Alcohol + acids

    phosphates ( glucose -1-phosphate)

     

    c.

    Dehydratin( Molisch, Selivanoffs, Bials tests)

    Strong acids (Con.H2SO4)

    Pentose→ Furfurals,Hexose→ Hydroxy methyl Furfural.

    TESTS

     

    Name of test

    Substrate

    Reagents

    Result

    1

    Molisch's Test

    Sugar

    3d Alfa-Naphthol in Methanol +H2SO4 (side)

    Violet color in the Junction.

    2

    Iodine test

    Starch

    Dil. Iodine

    Deep Blue Colour

    3

    Benedict's

    Reducing Sugar(0.5 ml/ 8 dps)

    CuSO4+Na2CO3+NaCitrate(5 ml)

    Cuprous Oxide (Red Precipitate)

    4

    Fehling's Test

    Reducing Sugar

    CuSO4+KOH+NaK Tartrate

    Cuprous Oxide (Red Precipitate)

    5

    Barfoed's test

    Monosaccharide.

    CuAcetate+Dil.AceticAcid

    (Red Precipitate) in < 5'

    6

    Bial's Test

    Pentose

    Orcinol Hydrochloride (Boil)

    Green

    7

    SelivanoffsTest

    Ketoses

    Dil. Resorcinol in Dil.HCl (heat)

    Cherry red

    8

    Tollent's Test

    Reducing Sugar

    Ammoniacal Silver nitrate

    Metalic Siver Mirror
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