Australian Regenerative Medicine Institute

Essential Role of Platelet-Derived: MATERIALS AND METHODS(2)

Tissue Preparation

To obtain tissue specimens, pregnant mice were killed by cervical dislocation, and intact uteri were removed. For both immunohistochemistry and in situ hybridization, embryos at 8.5 to 11.5 dpc were fixed with the uterine wall, whereas for embryos older than 12.5 dpc, the placental discs with yolk sacs were separated from the uterus and fetal bodies and fixed. The fixation procedure was previously described. After fixation, the tissues were dehydrated by passage through 70%, 85%, and 100% ethanol, and stored in methanol at — 20°C.

Immunohistochemistry

Fixed and dehydrated tissue specimens were embedded in polyester wax and sectioned at 6-8 |xm. Sectioned specimens were subjected to immunohistochemistry as previously described.

Whole-mount immunohistochemistry was performed as previously described except that the tissue blocks were embedded in 3% agarose gel and sliced at 400-500 (Jim using a Vibratome Series 1000 Sectioning System (Technical Products International). buy ortho tri-cyclen online

For immunohistochemistry, we used two established monoclonal antibodies as primary antibody—rat monoclonal anti-mouse PDGFRct antibody, APA5, and rat monoclonal anti-c-kit antibody, ACK2. We used horse radish peroxidase (HRP)-conjugated goat polyclonal anti-rat immunoglobulin antibody (Biosource, Camarillo, CA) as secondary antibody.

Category: Platelet-Derived

Tags: Growth Factor, Mouse Placenta, Platelet-Derived, Yolk Sac

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