Wallac 1470 wizard gamma

Aβ transport studies were performed as we previously reported (Qosa et al., 2014a (link); Qosa et al., 2014b (link)). bEnd3 cells were plated on 0.45 μm polyester membrane transwell 24-well plates, then treated with the compounds (10 μM) on day five for 24 h. Next day, Aβ₄₀ transport assay was initiated by addition of a mixture of 0.1 nM ¹²⁵I-Aβ₄₀ and 0.05 mM ¹⁴C-inulin in media to the B-side. At the end of incubation period, 30 min, aliquots from both sides were separately collected for radioactivity analysis using Wallac 1470 Wizard Gamma and Wallac 1414 WinSpectral Liquid Scintillation Beta Counter (PerkinElmer; Waltham, MA). The transport quotients of ¹²⁵I-Aβ₄₀ (TQ_B→A) was calculated using the following equation (Qosa et al., 2014a (link); Qosa et al., 2014b (link)):
$$A{\beta }_{\mathit{40}}TQ=\frac{(A{\beta }_{\mathit{40}(A)})/(A{\beta }_{\mathit{40}(B)})}{({\mathit{Inulin}}_{(A)})/({\mathit{Inulin}}_{(B)})}$$ where Aβ₄₀ (A) is the cpm of ¹²⁵I-Aβ₄₀ in the upper side, and Aβ₄₀ (B) is the cpm of ¹²⁵I-Aβ₄₀ in the lower side, inulin (A) is the dpm of ¹⁴C-inulin in the upper side, and inulin (B) is the dpm of ¹⁴C-inulin in the lower side. Three independent experiments were carried out for each compound.
Concentration-dependent studies for Aβ transport across the monolayer was performed for Tph in the concentration range between 1–10 μM as described above.

bEnd3 cellscultured on 24 well inserts were treated for 48 h with 2.2 μg/mL of Crocus sativus extract or vehicle. Aβ transport was performed as described previously.^{53 (link)} Briefly, treatments were removed and replaced with fresh media for 4 h, as a washout period. The basolateral to apical transport (B → A) was initiated by the addition of media containing 0.1 nM ¹²⁵I-Aβ₄₀ and ¹⁴C-inulin (0.1 μCi/mL) to B side (lower chamber) and media alone to A side (upper chamber) for 30 min. Aliquots from both A and B sides were taken for radioactivity analysis using Wallac1470 Wizard Gamma and Wallac 1414 WinSpectral liquid scintillation counters (PerkinElmer Inc. Waltham, MA).