LIST OF PROTOCOLS - P.J. HANSEN LAB 

Assays |  Embryos |  Immunochemical Procedures  |  Molecular Biology  |  Tissue and Cell Culture
Other Procedures  |  Other Links

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 ASSAYS

          Glutathione - a microassay for determining glutathione content in cells

Bradford Protein Assay  - a microassay for determining protein content in a 96-well microtiter plate format

Lowry Protein Assay

        EMBRYOS   

In Vitro Production of Bovine Embryos

Other protocols for embryos are listed below:

Apoptosis

FAM caspase 8 and 9 binding assay for embryos

Group II caspase activity in bovine embryos

TUNEL procedure for preimplantation embryos -  see Tutorial ImageJ for Luciano Bonilla's guide for using ImageJ for counting cell number. 

Cell Counting

Cell number counting in preimplantation embryos

 

Live/dead stain for embryos and oocytes

 

Differential staining of trophectoderm and inner cell mass of bovine embryos - see Tutorial ImageJ for Luciano Bonilla's guide for using ImageJ for counting cell number. 

 

Immunolocalization of CDX2

 Cryopreservation: Slow Freezing | Vitrification (Seidel) |  Open pulled straw vitrification (Vajta)  |  Open pulled straw vitrification in Spanish

 

Other

Detection of reactive oxygen species in preimplantation bovine embryos

Determining lipid content in embryos using Nile Red fluorescence

Immunodetection of 5-methylcytosine in preimplantation embryos

 

Immunolocalization of CDX2

 

Isolation of trophectoderm and inner cell mass from bovine blastocysts using magnetic activated cell sorting

Mini-Chamber for regulating gaseous environment during culture
 

Preparation of in-vitro produced bovine embryos for transfer into recipients

Searching for oocytes - the Movie! 

 

Sexing Bovine Preimplantation Embryos by PCR

 

Zona pellucida removal

        IMMUNOCHEMICAL PROCEDURES

Immunolocalization of CDX2

 

Immunoprecipitation

 

Semi-dry transfer of proteins for Western blotting

 

Western blotting using ECL reagent

       

        MOLECULAR BIOLOGY

Polymerase Chain Reaction - a detailed guide by Justin Fear

PCR using the Bio Rad CFX96 - a practical guide by Kyle Dobbs

RNA purification using TRizol    

        TISSUE AND CELL CULTURE

Culture of endometrial explants
 

Culture of BEND cells (a bovine endometrial cell line)  

 

Establishment of bovine fibroblasts from fetal skin

 

Hemacytometer, Use of

      OTHER PROCEDURES

          Acrosomal stain

      OTHER LINKS

Apoptosis Protocols – part of the Immunohistochemistry World Website     

Buffer Design  For those who have trouble designing buffers and calculating molarity, here is the site for you (from Professor Rob Benyon at University of Manchester

 

Crown rump length - use this site from John Parrish to calculate stage of gestation in cattle

Fixation Procedures for Flow Cytometry – From the University of Iowa, these work for immunocytochemistry too.

          Histotechnology (Univ. Nottingham)

          Immunohistochemistry Protocols from Immunohistochemistry World   

          Immunoportal - everything immunohistochemical

Molecular Biology Protocols 

Online Resources for Immunology Research Using Comparative and Veterinary Animal Models

Open Wetware - From MIT, the site "is an effort to promote the sharing of information, know-how, and wisdom among researchers and groups who are working in biology & biological engineering". 

Ovine Formulary  A list of protocols for inducing anesthesia in sheep

 

Power Calculations - from R.V. Lenth, University of Iowa

Protocols Online  An excellent source of protocols for biological research

University of Florida Flow Cytometry Core Lab – protocols, software and links

UNC Tissue Culture Facility - a great site, includes some tips on recognizing and dealing with contamination

Understanding and Managing Cell Culture Contamination - from Corning

University of Pennsylvania School of Dental Medicine Flow Cytometry Facility

WWW Virtual Library of Cell Biology - Methods, Protocols, and Software A good collection of links to lab protocols