Transam nfatc1 kit

NFATc1 and c2 transcription factor activity assays were performed using nuclear lysates prepared from primary mouse microglia cultures. Cells were plated in serum free DMEM/F12 media overnight before stimulation with 10 μMAβ_1–42 for 1 h at 37°C. After the stimulation was over, the plates were washed with PBS and nuclear lysates were prepared following the manufacturer protocol (Active Motif, Carlsbad, CA). The amount of protein in the nuclear lysates was determined by the Bradford assay [54 (link)] then used to perform activity assessments using the TransAm NFATc1 kit (Active Motif, Carlsbad, CA) with slight modification of the manufacturer protocol to substitute with an anti-NFATc2 detection antibody for quantifying NFATc2 activity as well. Activities of both NFATc1 and c2 were measured with a plate reader (Biotek, Winooski, VT) at 450 nm with a reference filter at 630 nm.