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Equipment and applied methods P&ToR

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)