Hansen Lab Protocols

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

Apoptosis

• FAM caspase 8 and 9 binding assay for embryos

• Group II caspase activity in bovine embryos

• TUNEL procedure for preimplantation embryos

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
• Cryopreservation: Slow Freezing | Vitrification

In Vitro Production

• In Vitro Production of Bovine Embryos (note the link on the index at this site for other embryo protocols)

Other

• Detection of reactive oxygen species in preimplantation bovine embryos
• Mini-Chamber for regulating gaseous environment during culture

IMMUNOCHEMICAL PROCEDURES

• Antigen retrieval for immunohistochemistry with paraffin-embedded tissues

• Immunoprecipitation
• Western blotting using ECL reagent

MOLECULAR BIOLOGY

• RNA purification using TRizol    

TISSUE AND CELL CULTURE

• Culture of endometrial explants
• Culture of BEND cells (a bovine endometrial cell line)
• Hemacytometer, Use of

• Acrosomal stain
• pH Meter (Accumet X15):  Protocol for use in PJH lab  Manual
• Rectal palpation of cattle
• Scintillation spectrometry - Manual for Beckman LSC-6000
• Synthesis of glutathione ester