UCD DERC Experimental Methods & Lab Protocols
(.pdf files)
Animal (click here for full pdf version)
A1: Ear Tagging
A2: Tail Snip
A3: Micro Tail Bleed / Blood Glucose
A4: Retro-Orbital Blood Sampling
A5: Streptozotocin Induced Diabetes
A6: Implantation of Alzet Pumps
A7: Implantation of Slow Release Pellets
A8: Isolation of Mouse Pancreatice Islets
A9: Dithizone Staining of Islets
A10: Islet Transplantation
A11: Staging of Embryos
A12: Dissection of Young Pancreatic Rudiments from Mouse
Cell Culture (click here for full pdf version)
C1: Protocol for Cell Line Establishment from Embryonic Tissues
C2: Harvesting Cells with Trypsin/EDTA
Diagnostic (click here for full pdf version)
D1: HLA Typing
D2: Insulin Autoantibody Immunoprecipitation Assay
D3: Combined GAD-65/ICA-512BDC In Vitro Translation & Immunoprecipitation Assay
D4: Mouse ELISPOT Assay
D5: Insulin Release and Immunoassay
Histology (click here for full pdf version)
H1: Fixation Issues and Frozen Block Generation
H2: Fixing Cells with Fresh Paraformaldehyde
H3: Preparation of Frozen Sections
H4: Immunohistochemistry
H5: Zymed Staining Procedure
H6: Double & Triple Immunofluorescent Staining on Parafin or Frozen Sections
H7: TSA-Amplification During Double- and Triple- Immunofluorescence Staining
H8: Counterstaining Procedures
H9: b-Galactosidase and Alkaline Phosphatase Staining Protocols
H9a: IacZ Wholemount Staining
H9b: hPLAP Wholemount Staining
H10: LacZ and hPLAP Slide Staining
H11: General Materials for Histology
H12: DIG-RNA In Situ Hybridization (Frozen Sections)
H13: Lectins
H14: Whole-Mount Immunostaining
H15: Immunohistological Morphometry ImagePro Plus
H16: Immunocytochemistry of Cell Monolayers (Chamber Slides)
Immunology (click here for full pdf version)
I1: Guidelines for the Culture of T Cell Lines and Clones
I2: Lymph Node Proliferation Assays and Establishment of Antigen Specific T Cell Lines
I3: Density Gradient Separation of Antigen Activated T Cells from Lymph Node Populations
I4: Polyethylene Glycol (PEG) Mediated T Cell Fusion
I5: Transfer, Expansion, Testing and Cloning of T Cell Hybrids
I6: Freezing BW5147 and Hybrid T Cells for LN2 Storage
Molecular Biology (click here for full pdf version)
M1: Genomic DNA Isolation from Mouse Tail Samples
M2: Multiplex-RT-PCR
M3: Amplification of Genes for ISH Probe Generation
M4: Enzymatic manipulations of DNA, Plasmids
M5: Bio-Rad Protein Assay (BRADFORD METHOD)
M6: Intracellular Flow Cytometry Staining Protocol
M7: Pancreas Digestion Protocol
All Protocols (combined .pdf document with all above protocols)
The purpose of this website is to provide some basic information about the DERC cores including contact information and links to on-line services where they are available. If you have questions about the UCD DERC please contact us (email or phone: 303.724.6836).
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