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Postculture media were assayed for VEGF and PGE2 by ELISA.
The suspensions in organic media were assayed for catalytic activity, and structures were probed by FT-IR and CD measurements.
VEGF in mouse serum and VEGF in cell culture media were assayed using the mouse (R&D Systems) and human (R&D Systems VEGF Quantikine ELISAA kits according to the manufacturer's instructions.
The protein levels of OPN in conditioned media were assayed by immunoblot using a primary antibody from Santa Cruz Biotechnology Santaa Cruz, CA).
The test media were assayed six times for each treatment condition.
After centrifugation, media were assayed to determine the level of antigen.
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Conditioned media was assayed for type-II collagen degradation fragments (CTX-II) and an aggrecan epitope (CS-846) associated with new aggrecan synthesis.
The total sugar concentration in the media was assayed by a colorimetric method (Dubois et al. 1956) using a A145 kit (Jiancheng Biotech Co., Ltd. China), and then converted into the amount of soluble starch consumed by multiplying a factor of 1.11 (1 unit soluble starch forms 1.11 unit glucose).
Accumulation of nitrites in media was assayed by the standard Griess reaction.
The LDH release in the media was assayed using in vitro LDH release kit from Sigma by following manufacturer's protocol.
The concentration of GnRH in the media was assayed using an LH-releasing hormone enzyme immunoassay kit S-12177, PeninsuLaboratoriesies, LLC, San Carlos, CA) according to the manufacturer's instructions.
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