Adenovirus Protocols

• PLAQUE ASSAYS FOR ADENOVIRUS TITRATION
• Preparation of High Titer Adenovirus in P11 cells

Bacterial Protocols

• Bacterial Expression of GST-fusion Proteins
• Bacterial Expression of IRS-1 containing GST-fusion Proteins
• Denaturing purification of insoluble bacterial GST-fusion Proteins
• Preparation of LB and LB+ Amp Plates

DNA/RNA Isolation and Analysis

• Isolation of total RNA from mouse muscle
• PHENOL-CHLOROFORM EXTRACTION
• Large DNA Prep/CsCl
• Preparation of Herring Sperm DNA for Southern Blotting
• Probe DNA Preparation for Southern Blotting
• SDS Page
• Southern Blot Protocol
• Synthesis of Double-Stranded cDNA from Purified Poly(A) mRNA
• Oligos into Linkers

ERROR ANALYSIS

• Error Propagation

GENECHIP ANALYSIS

• ChipInfo
• dCHIP
• GO Surfer

GENOTYPING

• Multiple Genotyping Protocols

GENOTYPING MOUSE TAIL DNA

• SYSTEMIC IRS1 and IRS2
• SYSTEMIC JNK 123
• SYSTEMIC Insulin related receptor (IRR)
• SYSTEMIC PTP1B
• SYSTEMIC PDX1 KO and PDX1 transgene
• SYSTEMIC IGF1R
• SYSTEMIC IRS4
• SYSTEMIC IRS3
• RIP IRS2 Transgene
• FLOXED IRS2

Histology and Staining

• Immunocytochemistry for Paraffin-embedded Sections

Hormone Assays

• ELISA Protocol

Immunoprecipitation and Western Blotting

• ACTIVATION OF p70s6k and MAP KINASES
• GENERAL PROTOCOL FOR IMMUNO-
• IN VIVO PHOSPHORYLATION/IP
• SDSPAGE
• STAINGEL
• Tryptic digest of SDS-PAGE Purified proteins
• WESTERN BLOTTING PROTOCOL
• WESTBLOT 7/28/98
• FOXO1 WESTERN BLOT
• IMMUNOPRECIPITATION

Instrument Manuals

• Microbeta Workstation v4.0
• ImageQuant TL v2003.03
• Fragment User Guide
• ImageQuant Utilities

JAXS Frozen Mouse Sperm

Kinase and Phosphatase Assays

• ACTIVATION OF p70s6k and MAP KINASES:
• Akt/PKB kinase assay
• In Vitro MAP kinase assay 9/1998
• IN-VITRO KINASE ASSAY
• JNK Kinase Assay
• JNK Kinase Assay
• PROTOCOLS 1
• PROTOCOLS 1

Live Animal Protocols

• Glucose tolerance test and fasting insulin test protocol (updated)
• GTT / ITT Data form
• Insulin tolerance test (updated)
• DEXA scanning protocol

Mammalian Cell Culture and Manipulation

• 35 S Met/Cys label Fao cells
• General Outline of Lipogenesis Assay (DRAFT VERSION)
• Anchorage-Independent Growth Assay
• Mammalian Cell Transfection – Calcium Phosphate Method
• Crystal Violet Staining (for Focus Formation Assay)
• General Protocol for an Epo Binding Assay with 293 Cell Line
• Glucose Transport Assay
• GROWING CHO CELLS
• FAOGROW
• RATGROW
• 32DGROW
• \PROTOCOLS\HIS100
• METHIONINE LABEL/IP
• MITOGENIC ASSAY FOR 32-D CELLS
• PC12 cell protocols
• Preparation of Neo (G418) Stock:
• Radiolabeled Ligand Binding Assay on 32D Cells
• PROTOCOLS\
• VEGF release assay
• WEHICM

Molecular Biology Protocols

• WHITE LAB – SOUTHERN BLOTTING PROTOCOL
• PCRMUT1/27/93
• LASER CAPTURE DISSECTION

OpenLab

• Open lab Preferences Setup
• Create a MAC Account for OpenLab
• Connecti OpenLab/MAC to WHITELABS

PCR Protocols

• Kyle’s protocol for long PCR
• PCR GENOTYPING PROTOCOL
• IRS-1 and IRS-2 Genotyping (PCR)
• PCRMUT1/27/93
• PCR Genotyping of Mouse Tails for IRS-1 and IRS-2

RECOMBINANT PROTEIN FORM

Protein Protocols

• PROTOCOLS 1
• PROTOCOLS 1
• PROBING SH2 DOMAIN-CONTAINING
• PRODUCTION OF RECOMBINANT (BACULOVIRUS PRODUCED) IRS-1 PROTEIN
• ANTIBODY PURIFICATION

Silicon Genetics

• Manual Version 6.1
• Genet Version 4.1

Southern Blotting Protocols

• WHITE LAB – SOUTHERN BLOTTING PROTOCOL
• Preparation of Herring Sperm DNA for Southern Blotting
• Probe DNA Preparation for Southern Blotting
• Protocol for Genotyping by PCR
• SOUTHERN BLOTTING ON GENOMIC DNA
• SOUTHERN PROBE DNA PREPARATION
• WHITE LAB – SOUTHERN BLOTTING PROTOCOL
• WHITE LAB – SOUTHERN BLOTTING PROTOCOL

Typhoon

• Fluorescence Imaging With Typhoon
• ECL Plus Western Blot Detection
• Detection of Biomolecules Part 1
• Detection of Biomolecules Part 2