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Thursday, 31 May 2007
Freezing and Thawing Eukaryotic Cells
From Dr. Bart Frank Library, Arthritis and Immunology Program, OMRF
Freezing cells in liquid nitrogen
For T25 Flask
- Take off supernatant
- Trypsinization with 1ml 0.25% EDTA
- Give a hard Shock to the flask to remove all attached cells
- Add 10ml Media +10%FCS
- Pipette up and down to distribute cells throughout media (i.e. not clumped together)
- Add resuspended cells to sterile universal tube
- Spin down 1500rpm, 3 mins
- Take off media
- Resuspend pellet in 2ml FCS +10%DMSO
- Distribute in 500ul aliquots (0.5 to 2 Million cells/ml)
- Move cells to -80oC overnight in Mr. frosty box (filled with ethenol)
- Finally freeze cells in liquid N2
Unthawing
- Warm DMEM in waterbath
- Thaw cryovial at 37oC quickly until cells become molten.
Aliquot the 1ml of cells using a disposable pipette into 10ml fresh media in a TC flask
Cell Thawing/Freezing Protocol
From University of Chicago
Tissue Culture - Storage of Cell Lines
From Hancock Laboratory Methods
Bioinformatics: A Practical Guide to the Analysis of Genes and Proteins Andreas D. Baxevanis, B. F. Francis Ouellette
Bioinformatics: A Practical Guide to the Analysis of Genes and Proteins
Andreas D. Baxevanis, B. F. Francis Ouellette
Introductory Course in Immunology June 22 - 28, 2007
The University of Pennsylvania, Philadelphia, Pennsylvania Don’t miss the most comprehensive introduction to immunology available!
An intensive course taught by world-renowned teachers and researchers in immunology. This 37 ½ hour, two-part course provides a comprehensive overview of the basics of immunology for students new to the discipline or seeking more information to complement general biology or science training. Upon completion students will have the tools necessary to understand and discuss current topics and methods in immunology. Part I (June 23-25) is a comprehensive introduction to the basic principles of immunology and is suitable for students with a general biology background. Part II (June 26-28) is a lecture course covering major areas of immunology and will require an understanding of basic immunology. Parts I and II may be taken independently at the discretion of the student. Financial support for underrepresented minority scientists is available through the FASEB MARC program. Registration Deadline: June 1, 2007. Course Director: Terri M. Laufer, University of Pennsylvania School of Medicine Part I: June 22-25
· Terri M. Laufer, Univ. of Penn School of Med.
Welcome and introduction to the immune system
· Kathleen E. Sullivan, Children’s Hospital of Philadelphia Introduction to innate immunity
· Jennifer A. Punt, Haverford College Introduction to adaptive immunity
· Moshe J. Sadofsky, Albert Einstein College of Med. VDJ recombination
· John G. Monroe, Univ. of Penn School of Med.
B cell development
· Terri M. Laufer, Univ. of Penn School of Med. Introduction to the MHC
· Laurence C. Eisenlohr, Jefferson Medical College
Antigen processing and presentation
· Al Singer, NCI, NIH
T cell development
· John C. Cambier, National Jewish Center, Colorado
Univ. Health Science Center
Signaling in the immune system
· Laurence A. Turka, Univ. of Penn School of Med. Costimulation
· Michael P. Cancro, Univ. of Penn School of Med.
Lymphocyte homeostasis
· Marc K. Jenkins, Univ. of Minnesota, School of Med. The primary immune response: putting the pieces back together
Part II: June 26-28
· Andrew J. Caton, Wistar Institute
T and B cell tolerance
· Gail A. Bishop, Univ. of Iowa
Made for each other: T-B cell collaboration in the immune response
· Cathryn Nagler, Massachusetts General Hospital
Mucosal immunology
· Betty Diamond, Feinstein Institute for Medical Research, North Shore-LIJ Health System
Autoimmunity
· Arnold I. Levinson, Univ. of Penn School of Med.
IgE –mediated hypersensitivity
· David Artis, Univ. of Penn School of Med.
Immunity to helminth parasites: new insights into the regulation of type 2 cytokine responses
· George S. Yap, Brown Univ.
Immunity to intracellular pathogens: a tale of hide and seek
· Joel D. Ernst, NYU School of Med.
Immunity to bacterial infections
· Barney S. Graham, NIAID, NIH
Immunization against viral diseases
· Wayne W. Hancock, Children’s Hospital of Philadelphia
Transplantation
· Jordan S. Orange, Children’s Hospital of Philadelphia Primary immunodeficiency
· Andy Hurwitz, LMI, CCR, NCI
Tumor immunology
· Hugh Auchincloss, NIAID, NIH
Current issues in immunology
FOR INFORMATION, COURSE OUTLINES, AND REGISTRATION, VISIT: www.aai.org/Courses.htm For assistance in registering, contact aaioffice@aai.org or 301-634-7178. Financial support for underrepresented minority scientists is available through the FASEB MARC Program. Overseas applicants are advised to apply early for visas
Monday, 28 May 2007
Saturday, 26 May 2007
Cytokine Protocols (Methods in Molecular Biology)
Pub Date: October 2003
Pages: 248
ISBN: 0-89603-948-X
DOI: 10.1226/089603948X
Table of Contents
Large-Scale Generation of Plasmids that Express Type I Interferon
Peter Härle and Daniel J. J. Carr
Identification of trans-Acting Factors by Electrophoretic Mobility Shift Assay
Rebecca J. Chan, Min You, and Gen-Sheng Feng
Modulation of the Interferon-g Signal by Transfection of Cells with an
Antisense-RNA Expressing Vector
Leen Delrue and Marc De Ley
Competitive RT-PCR to Quantify Small Amounts of mRNA
Gabriela Aust
In Situ Hybridization for Cytokines in Human Tissue Biopsies
Emily Mathey, John Pollard, and Patricia Armati
Purification, Identification, and Synthesis of Chemokines
Patricia Menten, Anja Wuyts, and Jo Van Damme
Receptor Isolation and Characterization: From Protein to Gene
Daniela Novick and Menachem Rubinstein
Crystallization of Cytokine–Receptor Complexes
William J. Cook and Mark R. Walter
Biosensor Analysis of Receptor–Ligand Interactions
Sheng-Jiun Wu and Irwin Chaiken
Analysis of SH2 Ligands and Identification of Sites of Interaction
Manuel Baca
Assays for Antiviral Activity
Anthony Meager
Assays for Cytotoxicity
Anthony Meager
Assays for Chemotaxis
Anja Wuyts, Patricia Menten, Nancy Van Osselaer, and Jo Van Damme
In Vitro and In Vivo Assays for High-Molecular-Weight Pegylated Muteins of
Granulocyte Colony-Stimulating Factor
James F. Eliason, Nadine Tare, Sharon Bowen, Tomoaki Inoue, and Ikuro Horii
Development of a Mammalian Tet-On Expression Cell Line: Glucosylceramide
Synthase Regulates TNF-a-Induced Apoptosis
Yong-Yu Liu and Myles C. Cabot
Immunohistochemical Detection of Cytokines in Human Tissue Sections
Sandra Hulshof, Danny F. Dukers, Wim Vos, and Corline J. A. De Groot
Detection of Cytokine- and Chemokine-Expressing Cells at the
Single Cell Level
Karin Loré and Jan Andersson
Detection of Cytokine Signal Transduction “Cross-Talk†in Leukocyte
Activation
Howard R. Petty
Assay System for the Effect of Prostaglandin E2 in the Determination of
Polarized Cytokine Production
Etsushi Kuroda and Uki Yamashita
Phospholipid Signaling Protocols (Methods in Molecular Biology, volume 105)
Phospholipid Signaling Protocols (Methods in Molecular Biology, volume 105)
Author: Bird, Ian
Publisher: Humana Press
Publication Date: 1998-06
Number Of Pages: 380
Average Amazon Rating:
Book Id 49833
Download Details:
Methods in Molecular Biology, vol.105
Download
Password: econiches
Electro Microscopy Methods and Protocols (Methods in Molecular Biology)
- Publisher: Humana Press
- Number Of Pages: 283
- Publication Date: 1999-04-15
- Sales Rank: 1795694
- ISBN / ASIN: 0896036405
- EAN: 9780896036406
- Binding: Plastic Comb
- Manufacturer: Humana Press
- Studio: Humana Press
Pass: deadzoners.tk
Introductory Plant Biology, By Kingsley R Stern, Jim Bidlack & Shelley
Introductory Plant Biology, By Kingsley R Stern, Jim Bidlack, Shelley
Jansky, Kingsley Stern,
* Publisher: McGraw-Hill Science/Engineering/Math
* Number Of Pages: 640
* Publication Date: 2003-04-23
* Sales Rank: 444653
* ISBN / ASIN: 0072930381
* EAN: 9780072930382
* Binding: Hardcover
* Manufacturer: McGraw-Hill Science/Engineering/Math
* Studio: McGraw-Hill Science/Engineering/Math
Download
Part 1
Part 2
Part 3
Part 4
Part 5
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Mathematical Biology by J.D. Murray
Mathematical Biology I, By J.D. Murray
* Publisher: Springer
* Number Of Pages: 584
* Publication Date: 2004-06-17
*
Sales Rank: 137251
* ISBN / ASIN: 0387952233
* EAN: 9780387952239
* Binding: Hardcover
* Manufacturer: Springer
* Studio: Springe
pass: tFMATBIOI.rar
Calpain Methods and Protocols (Methods in Molecular Biology)
Friday, 25 May 2007
Online Reference Library
xreferplus
Thursday, 24 May 2007
Cell Biology Laboratory Protocols
Laboratory Manual is one of the best online lab manual available on web, written by William H. Heidcamp, Gustavus Adolphus College Saint Peter, Minnesota, USA.
Chapter 1: The Microscope
- Introduction
- 1.1 The Bright Field Microscope
- 1.2 Use of Oil Immersion (100x)
- 1.3 Measurements: Ocular and Stage Micrometers
- 1.4 Measuring Depth
- 1.5 Measuring Area
- 1.6 Measuring Volume
- 1.7 Measurement of Cell Organelles
- 1.8 Use of Darkfield Illumination
- 1.9 The Phase Contrast Microscope
- 1.10 The Inverted Phase Microscope
- 1.11 The Transmission Electron Microscope
- 1.12 Comparison of Electron Micrographs
- Endnotes
Chapter 2: Histochemistry
- Introduction
- 2.1 Selective Staining: Prepared Slides
- 2.2 Basophilia
- 2.3 Periodic Acid Schiff (PAS) Reaction
- 2.4 Methyl Green-Pyronin Staining of DNA and RNA
- 2.5 The Feulgen Procedure for DNA
- 2.6 Alkaline Phosphatase Localization
- 2.7 Immunofluorescent Localization of Tubulin
- 2.8 Autoradiography
- Endnotes
Chapter 3: Cell Fractionation
- Introduction
- 3.1 Sucrose Fractionation of Castor Bean
- 3.2 Sedimentation Densities
- 3.3 Equilibrium Density Gradient - Percoll
- 3.4 Low Speed Separaton of Cells
- 3.5 Separation of Cells by Velocity Sedimentation
- Endnotes
Chapter 4: Electrophoresis
- Introduction
- 4.1 Preparation of SDS-polyacrylamide gels (SDS-PAGE)
- 4.2 Separation of protein standards: SDS-PAGE
- 4.3 Coomasie blue staining of protein gels
- 4.4 Silver staining of gels
- 4.5 Documentation
- 4.6 Direct densitometry of gels
- 4.7 Separation of LDH isozymes from serum
- 4.8 Western blots
- Endnotes
Chapter 5: Enzymes
- Introduction
- 5.1 Extraction of tyrosinase
- 5.2 Preparation of standard curve
- 5.3 Enzyme concentration
- 5.4 Effects of ph
- 5.5 Effects of temperature
- 5.6 Computer simulation
- 5.7 Kinetic analysis
- 5.8 Determination of Km and Vmax
- 5.9 Addition of enzyme inhibitors
- 5.10 Protein concentration/enzyme activity
- 5.11 Advanced analysis of enzyme parameters
- Endnotes
Chapter 6: Membranes
- Introduction
- 6.1 Lipid solubility of membranes
- 6.2 Osmosis
- 6.3 Pinocytosis
- 6.4 Computer simulation of membrane function
- 6.5 Extraction of brain lipids
- 6.6 Formation of lipid bilayer
- 6.7 Electrical properties of artificial membranes
- 6.8 Measurement of transmembrane potential
- 6.9 Chemical composition of myelin
- 6.10 Myelin lipid extraction
- 6.11 Quantitative analysis of lipid classes by TLC
- Endnotes
Chapter 7: Microsomes
- Introduction
- 7.1 Lysosome isolation in isotonic sucrose
- 7.2 Em observations of microsomes
- 7.3 Chemical characterization - acid phosphatase
- 7.4 Chromaffin granules from adrenal glands
- 7.5 Measurement of catecholamines
- Endnotes
Chapter 8: Photosynthesis\Respiration
- Introduction
- 8.1 Isolation of Chloroplasts from Spinach Leaves
- 8.2 ChlorophyII Content
- 8.3 The Hill Reaction
- 8.4 Isolation of Mitochondria
- 8.5 Respiration: Succinic Dehydrogenase
- 8.6 Cytochrome Oxidase Manometric Analysis
- 8.7 Respiration of Mitochondria
- 8.8 Photosynthesis by Respirometry
- Endnotes
Chapter 9: Tubules\Filaments
- Introduction
- 9.1 Fluorescent Analysis of Microtubules
- 9.2 Actin & Myosin-Microscopic Observation of Muscle
- 9.3 Histology of Extracellular Matrix
- 9.4 Isolation of Microtubules (Bovine Brain)
- 9.5 Viscosity & Polymeriztion of Microtubules
- 9.6 Polymerization Induced Changes in Absorbance
- 9.7 TEM Visualization of Microtubules
- 9.8 SDS Gel Electrophoresis of Tubulin\MAPs
- 9.9 Isolation of Actin and Myosin Filaments
- 9.10 Further Studies
- Endnotes
Chapter 10: Chromosomes
- Introduction
- 10.1 Polytene Chromosomes of Dipterans
- 10.2 Salivary Gland Preparation (Squash tech.)
- 10.3 Extraction of Chromatin
- 10.4 Chromatin Electrophoresis
- 10.5 Extraction and Electrophoresis of Histones
- 10.6 Karyotype analysis
- 10.7 In Situ hybridization
- Endnotes
Chapter 11: Cell Cycles
- Introduction
- 11.1 Mitosis
- 11.2 Grasshopper Spermatogenesis
- 11.3 Ascaris Oogenesis
- 11.4 3H - Thymidine Uptake by Cultured Cells
- 11.5 Timing of Cycles
- 11.6 Vicia Faba and Chemical Damage
- Endnotes
Chapter 12: Cell Cultures
- Introduction
- 12.1 Aseptic Cell Transfers
- 12.2 Examination of Bacterial Colonies
- 12.3 Gram Stain (+\-)
- 12.4 Prokaryote Cell Number by Dilution Plating
- 12.5 Cell mass by Measurement of Turbidity
- 12.6 Transfer of Eukaryote Suspension Cultures
- 12.7 Viability Cell Count
- 12.8 Computation of Transfer Aliquots
- 12.9 Eukaryote Growth Dynamics
- 12.10 Establishment of a Primary Culture
- Endnotes
Chapter 13: Differentiation
- Introduction
- 13.1 Sea urchin embryology
- 13.2 Determinants and fate - C. elegans embryos
- 13.3 The hematopoietic system
- 13.4 Photomorphogenesis
- 13.5 D.discoideum growth on agar
- 13.6 Suspension cultures of D. discoideum
- 13.7 Isolation of fruiting bodies without slugs
- 13.8 Prestalk vs prespore effect
- Endnotes
Chapter 14: Nucleic Acids
- Introduction
- 14.1 Extraction of DNA from bovine spleen
- 14.2 Purification of DNA
- 14.3 Characterization of DNA
- 14.4 DNA - dische diphenylamine determination
- 14.5 Melting point determination
- 14.6 CsCl - density separation of DNA
- 14.7 Phenol extraction of rRNA (rat liver)
- 14.8 Spectrophotometric analysis of RNA
- 14.9 Orcinol determination of RNA
- 14.10 Sucrose density fractionation
- 14.11 Nucleotide composition of RNA
- 14.12 Other studies
- Endnotes
Chapter 15: The Central Dogma
- Introduction
- 15.1 Protein Synthesis In Cell Free Systems
- Endnotes
Appendices
- Appendix A: Units and Measures
- Appendix B: Statistics
- Appendix C: Graphs
- Appendix D: Computers
- Appendix E: Image Analysis
- Appendix F: Centrifugation
- Appendix G: Spectrophotometry
- Appendix H: Radioactive tracers
- Appendix I: Photography
- Appendix J: Chemical Preparations
* Table of contents are adopted from
http://homepages.gac.edu/~cellab/index-1.html.
Gene pridiction software sites
- HMMgene - Prediction of genes in vertebrate and C. elegans
- Gene Discovery Page
- FramePlot - protein-coding region prediction tool for high GC-content bacteria
- tRNAscan-SE Search for transfer RNA genes in genomic sequence
- NETGENE - Predict splice sites in human genes
- ORF Finder
- BCM Gene Finder
- Grail
- Genemark
- Genie: A Gene Finder Based on Generalized Hidden Markov Models
- GENSCAN - predict complete gene structures
- Splice Site Prediction by Neural Network
- Procrustes
- GenePrimer
- GenLang
- MZEF
Gene Finder - Webgene - Tools for prediction and analysis of protein-coding gene structure
- MAR-Finder - Nuclear matrix attachment region prediction
- Glimmer bacterial/archael gene finder
Translate DNA/RNA to Protein
Translation tool from ExPASy
Translate is a tool which allows the translation (6 frames) of a
nucleotide (DNA/RNA) sequence to a protein sequence.
Wednesday, 23 May 2007
http://lectures.molgen.mpg.de/online_lectures.html
Learn Bioinformatics online
Max Planck Institute for Molecular Genetics provides a extensive online course for learning bioinformatics online.
Tuesday, 22 May 2007
List of companies selling Antibodies
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