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Example sentences for: temecula
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The blots were first blocked with 5% non-fat dry milk in TPBS buffer (PBS with 0.5% Tween 20) at RT for 1 hr, and then incubated with 1:3000 dilution of anti-β-actin (Sigma, St. Louis MO), 0.75 μg/ml of anti-NR1 (Chemicon, Temecula, CA), 1:1000 dilution of anti-NR2A (Chemicon), 0.35 μg/ml of anti-NR2B (Chemicon) and 1 μg/ml of anti-GluR1 (Chemicon) antibodies, respectively, at 4°C overnight.
Primary antibodies used: mouse anti-chicken Coll I, Coll III, FN, and TN (Hybridoma Bank, Iowa), rabbit anti-chicken TN (Chemicon, Temecula CA).
Collagen and elastin deposition in cell and matrix layers was determined by solid-phase ELISA with antibodies against goat anti-collagen I (Chemicon, Temecula, CA, USA), mouse anti-collagen III (Telios, San Diego, CA, USA), mouse anti-collagen VI (Telios), or mouse anti-elastin (Sigma), at dilutions recommended by the manufacturers.
Blots were washed twice with TBS-T containing 0.5% milk, then incubated at room temperature for 2 hours in TBS-T containing 0.5% milk and 1:5000 dilutions of secondary Ab conjugated to peroxidase (for BMP-5, anti-goat IgG-peroxidase from Chemicon, Temecula CA; for Smad1, anti-rabbit Ig-peroxidase from Amersham, Piscataway, NJ).
Polyclonal antibodies used were MyoD (Santa Cruz Biotechnology, Santa Cruz, California, United States) and nestin (gift from R. McKay); monoclonal antibodies were vimentin, alpha smooth muscle actin, fast-switch myosin, pan-cytokeratin (all from Sigma), and human nuclear antigen (Chemicon, Temecula, California, United States).
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