CTNNB1 monoclonal antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store at -20°C. -Aliquot to avoid repeated freezing and thawing.	-Mouse (1:2000)

Upstream tips
-Store at -20°C. -Aliquot to avoid repeated freezing and thawing.
Protocol tips
-Mouse (1:2000)

Publication protocol

Protein from MCF7 and MDA-MB-231 parental or mammosphere-derived cells pre-exposed to different treatments, namely, anti-CDH11 antibody, miR-335 mimic, or miR-335 inhibitor, were loaded (20 μg/lane) and separated using 10% sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE), then the blots transferred onto polyvinylidene difluoride (PVDF) membranes, blocked in 5% skim milk in TRIS buffered saline (TBS) with Tween-20 (TBST) for 1 h, at room temperature, then incubated overnight at 4 °C with the primary antibodies against CDH11 (1:1000, clone 2C67, #H00001009-M25), E-cadherin (1:1000, clone 7H12, #MAB16359), vimentin (1:1000, clone 1G3, #H00007431-M10), b-catenin (1:2000, #MAB11143), FYN (1:1000, clone 3G11-F9, #H00002534-M01), EZH2 (1:1000, clone 2C3, #H00002146-M01), and b-actin (1:2000, clone 3G4-F9, #H00000060-M01) all purchased from Abnova (Abnova Corporation, Taipei, Taiwan). This was followed by incubation in peroxidase - conjugated secondary antibody for 1 h at room temperature, and washed with TBST three times. The signals were developed using enhanced chemiluminescence (ECL-Plus, Amersham Pharmacia Biotech, Piscataway, NJ, USA) and detected with a UVP BioSpectrum® Imaging System (Analytik Jena US LLC, Upland, CA, USA).

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Manufacturer protocol

Download the product protocol from Abnova for CTNNB1 monoclonal antibody below.

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