ampullae excised into warm (37 °C) sterile EmbryoMax M-2 culture medium (EMD Millipore, Billerica, MA). Cumulus cells were removed with 0.5% hyaluronidase and mechanical pipetting. Washed, stripped oocytes were pooled in 100-μl droplets of M-2 under warm mineral oil (Irvine ...
To detect hypoxic areas in brain, rats were injected intraperitoneally with hypoxyprobeTM-1 (60 mg kg−1, solid pimonidazole hydrochloride; Natural Pharmacia International, Burlington, MA, USA) at 3 h before being killed. Isolated brains were immediately immersed in 4% paraformaldehyde for ...
cut-off of 5 kDa (Millipore, Billerica, MA). After ATP removal, a solution of 10 μM NBD1 in 20 mM Tris-Cl (pH 7.4), 2 mM TCEP, 2 mM MgCl2 was prepared. Aliquots of this solution were equilibrated with 0–20 µM TNP-ATP (Sigma) in the absence or presence 0....
After filtration, the digestion products were adjusted to 100 mL volume with Milli-Q water (18.2 MΩ, Milli-Q Element A10 purification system, Millipore, Billerica, MA, USA). Extracts were analyzed for TE concentrations determined by the Atomic Absorption Spectrometry (AAS) method using an iCE...