Preparation of the material for the studies:
- The isolation of cells of corpus luteum, myometrium, endometrium and epithelium of oviduct
- The cell cultures and strip incubation: an incubator (Memmert, Germany)
Analytical methods:
- Viability of cells determination (cytotoxicity test): ELISA (Multiscan EX, Labsystems, Finland)
- Enzyme immunoassay of steroids, protein hormones and prostaglandins (ELISA, Multiscan EX, Labsystems, Finland)
- PCR and its modifications: RT-PCR, nested-PCR, touchdown PCR
- RT PCR and Real-time PCR:
- RNA isolation (spectrophotometer BioPhotometr, Eppendorf, and Nonodrop)
- Reverse transcription, PCR (termocycler MJ Mini, BioRad)
- Real-time PCR (access to: ABI Prism 7300, Applied Biosystems, USA)
- Gel documentation system for electrophoresis gels (MiniBIS Pro, DNR Bio Imaging System, Israel)
- 5’ and 3’ RACE PCR – Rapid Amplification of cDNA Ends PCR
- Western Blot analysis (Mini PROTEAN 3 System)
- Intracellular Ca2+ mobilization using Fura 2 (microscope Nikon Eclipse TS 100 NIS-Elements Basic Research software)
- Immunohistochemistry (access to: microscope Imager Z1-Zeiss, Axio Vision 4.8 software)
- Measurement of contractility of myometrial and oviductal strips: isometric contraction transducer (access to: the HSE Type 372, Hugo Sachs Elektronik, Germany)
- Determination of catecholamine concentrations by HPLC with electrochemical detection (Hewlett-Packard, Series 1050)
