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(Restricted Access)
IMPORTANT NOTE
If you notice any errors in the information documents below or if you have found some improvement to the procedure, please immediately notify Gelb so he can update the document on the web.
Must-Read Handouts
(all Gelb group members are required to read these handouts before beginning research)
Lab Protocols
Biochemistry
- Arachidonate/PAF Analysis
- cPLA2-Alpha Purification
- Blurry Bands on Laemmli Gels
- 3H-DPPC/POPG sPLA2 Assay
- Eicosanoid LC-MS/MS
- Extinction Coefficient of Proteins at 280 nm Calculation
- Fatty Acid Binding Protein Purification and PLA2 Assay
- Fluorography of Gels
- Hamilton Syringes
- Handling Bee Venom PLA2
- HPLC Column Selection Guide
- Varian 210 HPLC Method Cheat Sheets
- Laemmli Gel of Total Ecoli Protein
- Lyophilizing Samples
- Lysophospholipid mass spec analysis
- Maleimide Probes
- Palmitoyl-Pyrenedecanoyl-phosphatidylglycerol and methanol PLA2 Assays
- Phospholipid Mass Spec Tables
- Phosphorous Analysis
- Precipitating Proteins for Laemmli Gels
- Removing SDS from Proteins
- Refolding sPLA2s in vitro
- Silver Staining Gels
- TRFIA
- Tris-Tricine Protein/Peptide Gels
- Urea
- PLA2-Vesicle Binding Assay Using Sucrose-Loaded LUVs
- PLA2 Assay with Radiolabeled E. coli Membranes
- Western Blots with ECL Detection
- Western Blot Analysis of Human sPLA2s
Molecular Biology
- Baculovirus
- Chemically Competent Bacteria
- DNA Quantification with Fluorescent Hoechst Stain
- DNA Sequencing (ABI PRISM Big-Dye Terminator Kit and ABI PRISM 310 Sequencer)
- Electrocompetent Bacteria
- Electroporation
- Fluorescence Microscope Lamp Changing (Gelb Lab)
- Genotyping Mice
- Immunofluorescence Mammalian Cells
- Immunofluorescence HEK293 Cells
- Microinjection (Gelb Lab and Keck Center)
- PCR and Southern Blot Analysis of ES Cell Clones
- RNA Interference
- RNA Isolation and Analysis RT-PCR (semi-quantitative)
- Site-Directed Mutagenesis (QuickChange and Sculptor)
- Strain Collection
- Transfection of Mammalian Cells
Synthetic Chemistry
- Cold Baths
- Diazomethane Generation
- Evans pKa Tables
- Flash Chromatography Handling Air-Sensitive Reagents
- Isco Flash System
- ISIS Mentor Databases including ACD
- NMR Solvent Chemical Shifts
- 3H-Phosphatidylglycerol Synthesis SciFinder User Guide (MacIntosh)
- Solvent Dispensing System Solvent Drying
- TLC stains
Miscellaneous Information
- Checking In to the Gelb Lab
- Checking Out of the Gelb Lab
- Strain Collection Forms (Microorganisms and Animal Cells)
- Tissue Culture
- Cambridge Crystallographic Database (substructure searching and retrieving coordinates)
- Padlock Combinations
- PyMol User Guide
Catalogues and Directories
- Biochemistry UW Stockroom Catalog
- BioSupplyNet Catalog of Biomedical Reagents from Numerous Companies ChemSources Online