Anti-Collagen III antibody (ab7778)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Rabbit (1:8000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Rabbit (1:8000) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

Western blot analysis was performed to detect the protein expression levels. In brief, tissues and cells were solubilized in cold RIPA lysis buffer. Proteins were separated by 10% SDS-PAGE, and transferred onto PVDF membranes, which were then incubated with PBS containing 5% milk overnight at 4°C. The PVDF membranes were then incubated with primary antibodies [collagen I, (2 µg/ml, ab6308); collagen III (1:8,000, ab7778); FN, (3 mg/ml, ab6328); MMP13 (1 µg/ml, ab84594); TIMP-1 (0.2 µg/ml, ab126847); TIMP-2 (1:1,000, ab1828); all from Abcam] at room temperature for 3 h, respectively, and then secondary antibodies [goat anti-mouse (ab6785, 1:10,000); goat anti-rabbit (ab6721, 1:20,000); donkey anti-goat (ab6881, 1:10,000); all from Abcam] at room temperature for 1 h. An ECL kit was then used to perform chemiluminent detection. Relative protein expression was analyzed using Image-Pro Plus software 6.0, represented as the density ratio vs. GAPDH.

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Manufacturer protocol

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