Rabbit anti ntrk2

Paraffin sections were cut at a thickness of 4 μm for mice (n = 31), rats (n = 11), humans (n = 10), pigs (n = 3), and horses (n = 2). Sections were separately stained for BDNF and Ntrk2 using a 1∶200 dilution of rabbit anti-BDNF (Abcam) or rabbit anti-Ntrk2 (Abcam), as above. Negative sections were incubated with normal goat serum in lieu of primary antibody. Images were captured by an Infinity camera (Lumenera Corp.) under 200X magnification on an Olympus IX81 microscope. Bat sections (n = 6) were stained at the State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, in Beijing, China using anti-BDNF (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and anti-Ntrk2 (Santa Cruz) antibodies as above.

Extracted protein (60 μg) from human endometrium was run on a 4–20% gradient gel (Thermo-Scientific) at 150 V for 50 minutes. Protein was transferred to PVDF membrane (VWR International, Mississauga, ON, Canada) at 40 V for 90 minutes. Blots were blocked for 1 hour at room temperature with 5% skim milk/TBS-T, and subsequently probed with 1∶1000 rabbit anti-BDNF (Abcam) or 1∶1000 rabbit anti-Ntrk2 (Abcam), overnight at 4°C. Anti-Rabbit-ECL secondary (GE, Mississauga, ON, Canada) at a concentration of 1∶5000 was applied for 1 hour at room temperature, blots were briefly washed in TBS-T and TBS, then incubated with ECL substrate (Thermo-Scientific) for 5 minutes. Exposures were performed using x-ray film (Thermo-Scientific), and the exposure times were 60, and 45 minutes for BDNF and Ntrk2 respectively.