Cells were harvested and lysed in nondenaturing lysis buffer (Applygen Technologies, Beijing, China). Equal quantities (40 µg of protein) of cell extract were resolved by 10% SDS–PAGE, and the resolved proteins were electrophoretically transferred to PVDF membrane and blocked with 5% fat-free dry milk in TBST for 2 h at room temperature. Then respective primary antibodies were added in 5% milk TBST and incubated overnight at 4°C and incubated with horseradish peroxidase-conjugated secondary antibodies for 2 h at room temperature and washed three times before detection. The blot was developed with LAS4000 enhanced chemiluminescence system (GE Healthcare, Buckinghamshire, UK) and the densities of the bands were determined using Gel-Pro Analyzer 4.0 software.
Las4000 enhanced chemiluminescence system
Manufactured by GE Healthcare
Sourced in United States
The LAS4000 enhanced chemiluminescence system is a high-performance imaging device designed for analyzing chemiluminescent signals. It offers advanced features for capturing and quantifying luminescent data from a variety of samples, including Western blots, ELISA, and other chemiluminescence-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using las4000 enhanced chemiluminescence system
1
Western Blot Protein Analysis
2
Western Blot Analysis of Estrogen Receptors
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The membranes were incubated with 1:1000 dilutions of primary rabbit anti-estrogen receptor alpha (ab3575, Abcam, Cambridge, UK), rabbit anti-estrogen receptor β (ab3576, Abcam, Cambridge, UK) and anti-GAPDH (1:50000, 60004-1-lg, Proteintech, Rosemont, USA) antibodies at 4 °C overnight. The next day, after washing with PBS three times for 10 min each, the membranes were incubated with the secondary antibodies for 1 h at room temperature. Finally, the target proteins were visualized using a LAS4000 enhanced chemiluminescence system (GE Healthcare, Wisconsin, USA).
3
Western Blot Analysis of Estrogen Receptors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The membranes were incubated with 1:1000 dilutions of primary rabbit anti-estrogen receptor alpha (ab3575, Abcam, Cambridge, UK), rabbit anti-estrogen receptor β (ab3576, Abcam, Cambridge, UK) and anti-GAPDH (1:50000, 60004-1-lg, Proteintech, Rosemont, USA) antibodies at 4 °C overnight. The next day, after washing with PBS three times for 10 min each, the membranes were incubated with the secondary antibodies for 1 h at room temperature. Finally, the target proteins were visualized using a LAS4000 enhanced chemiluminescence system (GE Healthcare, Wisconsin, USA).
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