Subsequent to 24 h, the treated and untreated cells were harvested and lysed, and the supernatants were separated from the cell debris by centrifugation at 13,000 × g for 15 min at 4°C. Aliquots containing 30 μg of total protein were separated by SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were then probed with primary rabbit monoclonal antibodies against MMP-2 and -9 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) at 4°C overnight. The membranes were subsequently probed with a goat anti-rabbit secondary antibody conjugated with horseradish peroxidase (Santa Cruz Biotechnology, Inc.) and visualized by electrochemiluminescence. Protein band densities were quantified using Bio-Rad Quantity One software (Bio-Rad, Hercules, CA, USA).
Goat anti rabbit secondary antibody conjugated with horseradish peroxidase
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Goat anti-rabbit secondary antibody conjugated with horseradish peroxidase is a laboratory reagent used in various immunoassay techniques. It functions as a detection agent, binding to and labeling rabbit primary antibodies with the enzyme horseradish peroxidase.
Automatically generated - may contain errors
2 protocols using goat anti rabbit secondary antibody conjugated with horseradish peroxidase
1
Quantification of MMP-2 and MMP-9 Protein Levels
2
Lung Tissue Protein Isolation and Analysis
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Cytoplasmic protein was isolated from lung tissues as previously described with modifications [24] . Samples were separated on a 10% SDS-polyacrylamide gel, transferred to PVDF membrane and probed with the rabbit anti-Hsp90 and anti-GR (Santa Cruz, USA) primary antibodies. The samples were then incubated with goat anti-rabbit secondary antibody conjugated with horseradish peroxidase (Santa Cruz, USA) and detected with an ECL substrate kit (Thermo Scientific, USA). In these experiments, β-actin (Santa Cruz, USA) served as an endogenous control. The intensity of the bands was analyzed by a UVP Bio-imaging acquisition and image analysis system (Ultra-Violet Products Ltd, UK).
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