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Then, the buffer was collected for NO measurement using the DAN method.
The wash buffer was collected for protein quantification using indirect ELISA.
An EDTA sample with 'magic buffer' was collected for genomic analysis (7D only).
100 mM NaCl/50 mM Tris HCl buffer was collected following the same procedures to obtain fraction NaCl-2. 1 M NaCl/50 mM Tris HCl buffer was collected for the last elution to obtain fraction NaCl-3.
The cells were washed and incubated with control or NMDA in the above buffer for 1 h, after which, the buffer was collected for NO measurement using the DAN method.
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After 24 hours, whole-cell lysates prepared with passive lysis buffer were collected for reporter detection by the Dual Luciferase Reporter System (Promega) according to the manufacturer's protocol.
The protein was dialyzed extensively against the same buffer used for NMR, and the background diffraction from the buffer was collected and subtracted.
A blank buffer spectrum was collected for each buffering system and subtracted from the results prior to analysis.
After the flow through, the column was washed three times using 500 ml of MACS buffer and the eluted cell fraction was collected for further experiments.
eNOS was then eluted from the bead antibody eNOS complex using the loading buffer, and the supernatant containing the eNOS was collected for SDS PAGE.
Cells were lysed in 1X passive lysis buffer in 48 h and the supernatant was collected for Dual-luciferase activity measurement (Promega, Madison, WI).
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