Anti-MMP9 antibody (ab73734)

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 (1:1000) -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 (1:1000) -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

Cell lysis solution was added into each group of cells. After 30 min dissociation at 4°C, the solutions were centrifuged at 11,670 g for 10 min. The supernatant was collected, which contained total protein. The protein concentration was measured by BCA kit (Beyotime). 20 µg protein in each group was loaded for gel electrophoresis and the membrane was transferred by wet methods. The antibodies, including anti-DEK (1:1,000, catalogue no. ab221545), anti-IMP3 (1:1,000, catalogue no. ab177477), anti-E-cadherin (1:1,000, catalogue no. ab1416), anti-Vimentin (1:1,000, catalogue no. ab8978; Abcam, Cambridge, UK), anti-MMP-9 (catalogue no. ab73734; 1:1,000; Abcam) and β-actin (1:1,000; catalogue no. AF0003, Beyotime Institute of Biotechnology, Shanghai, China) were incubated with nitrocellulose membranes overnight at 4°C. Next, the secondary antibody (1:100; catalogue nos. ab131368; Abcam) was added and co-incubated for 1–2 h at room temperature. ECL exposure liquid droplet (catalogue no. RPN2133; GE Healthcare Life Sciences, Chalfont, UK) was added on the membrane. In the end, the membrane was used for exposure utilizing gel imaging system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Grey density was analyzed using Quantity One analysis software v1.4.6 (Bio-Rad Laboratories, Inc.).

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Manufacturer protocol

Download the product protocol from Abcam for Anti-MMP9 antibody (ab73734) below.

Download manufacturer protocol

Videos

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