Cells were lysed in Laemmli buffer and the protein extracts were tested by Ab specific for OPRS1/Sigmar-1 (Abcam, Cambridge, UK), and β-actin (Sigma) diluted at 1∶500 and 1∶000, respectively. Anti-rabbit Ab conjugated to horseradish peroxidase (GE Healthcare, Little Chalfont Buckinghamshire, UK) was used as the secondary Ab at a dilution of 1∶5000. The SuperSignal enhanced chemiluminescence system was used for probing target proteins (Thermo Scientific, Rockford, IL). After the membranes had been probed for OPRS1/Sigmar-1, they were stripped and re-probed for β-actin.
Supersignal enhanced chemiluminescence system
Manufactured by Thermo Fisher Scientific
The SuperSignal enhanced chemiluminescence system is a laboratory equipment used for the detection and analysis of chemiluminescent signals. It provides a reliable and sensitive platform for the visualization and quantification of protein targets in Western blot applications.
Automatically generated - may contain errors
3 protocols using supersignal enhanced chemiluminescence system
1
Western Blot Analysis of OPRS1/Sigmar-1
2
Western Blot Analysis of 5-HT2B Receptor
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Cells were lysed in Laemmli buffer and the protein extracts were tested by Ab specific for 5-HT2B receptor (Abcam, Cambridge, UK), and β-actin (Sigma-Aldrich, St. Louis, MO, USA) diluted at 1:500 and 1:000, respectively. Anti-mouse Ab conjugated to horseradish peroxidase (GE Healthcare, Little Chalfont Buckinghamshire, UK) was used as the secondary Ab at a dilution of 1:5000. The SuperSignal enhanced chemiluminescence system was used for probing target proteins (Thermo Scientific, Rockford, IL). After the membranes had been probed for 5-HT2B, they were stripped and re-probed for β-actin.
3
Western Blot Analysis of Cell Signaling
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Cells were lysed in Laemmli buffer and the protein extracts were tested by Ab specific for Sig-1R/OPRS1, HIF-1α, ATF6, p65, phospho-p65 (S536) (all from Abcam), and β-actin (Sigma, Schnelldorf, Germany) diluted at 1:500 and 1:000, respectively. Anti-rabbit Ab conjugated to horseradish peroxidase (GE Healthcare, Little Chalfont Buckinghamshire, UK) was used as the secondary Ab at a dilution of 1:5000. The SuperSignal enhanced chemiluminescence system was used for probing target proteins (Thermo Scientific, Rockford, IL). After the membranes had been probed for Sig-1R/OPRS1 or HIF-1α, they were stripped and re-probed for β-actin.
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