cardiac LOXL1 and TGF-β1 were determined by immunoblotting. Briefly, proteins from the LV extracts were loaded in SDS-PAGE gels under non-reducing conditions and electrotransferred to a nitrocellulose membrane. Membranes were incubated with an anti-LOXL1 antibody (Abcam, Cambridge, MA) or with a mouse anti-TGF-β1 antibody (R&D Systems, Minneapolis, MN), then incubated with secondary antibodies conjugated with horseradish peroxidase (Cell Signaling Technology, Danvers, MA). Films were examined with a Perfection 3200 scanner (Epson America, Long Beach, CA) and band density and intensity quantified by densitometry. Increases in LOXL1 and TGF-β1 were normalized to GAPDH and expressed as fold increase vs control.
Mouse anti tgf β1 antibody
Manufactured by R&D Systems
Mouse anti-TGF-β1 antibody is a monoclonal antibody specific for the transforming growth factor beta 1 (TGF-β1) protein. It can be used for the detection and quantification of TGF-β1 in various experimental applications.
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2 protocols using mouse anti tgf β1 antibody
1
Cardiac LOXL1 and TGF-β1 Quantification
2
Quantification of TGF-β1 in Tenocyte Culture
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After VV treatment, 100 μl of the tenocyte culture medium was transferred to a 96-well ELISA plate that had been precoated with mouse anti-TGF-β1 antibody (R&D Systems, Minneapolis, MN), and the plate was incubated for 2 h at room temperature. After the wells had been washed 3 times with PBST, biotinylated anti-TGF-β1 antibody (R&D Systems) was added to each well, and the plate was incubated for 2 h at room temperature. After being washed to remove any unbound antibody-enzyme reagent, streptavidin horseradish peroxidase was added, and the plate was incubated for 20 min at room temperature in the dark. After washing, the substrate solution (a 1:1 mixture of H2O2 and tetramethylbenzidine) was added for color development, and then, the reaction was terminated by the addition of 2 N H2SO4. The absorbance of the plate was measured with a microplate reader at 450 nm (Dynex Technologies).
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