The HEY cells were treated with 2 at the concentration of 5 and 10 μM, respectively, 24 h later, they were splitted in RIPA lysis buffer (Beyotime, Jiangsu, China) including 1% PMSF (Thermo, Waltham, MA) at 4 °C for 30 min, then centrifuged at 10,000 rpm for 30 minutes. The concentration of protein was measured using a BCA Protein Assay Kit (Beyotime, Shanghai, China), then boiled for 10 minutes at 100 °C, the collected protein was carried out at 30 μg/per lane and separated on SDS-PAGE gel, electrophoretically transferred to a polyvinylidene difluoride membrane (Beyotime, Shanghai, China). The primary antibody incubated overnight and the secondary antibody incubated at 25 °C for 2 h, the bands were detected by super ELC detection reagent (Beyotime, Shanghai, China). Images were determined by Image-Pro Plus software (version 7.0, Olympus, Tokyo, Japan). All experiments were repeated 3 times.
Super elc detection reagent
Manufactured by Beyotime
Sourced in China
The Super ELC detection reagent is a laboratory product designed for use in electrochemiluminescence (ECL) detection applications. It serves as a core component in ECL-based assays, providing the necessary reagents to generate a luminescent signal that can be measured and quantified. The product description is limited to its core function without further interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Image pro plus software, by Olympus (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Polyvinylidene difluoride membrane, by Beyotime (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Thermo Fisher Scientific (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Lysate buffer, by Beyotime (1 mentions)
2 protocols using super elc detection reagent
1
Western Blot Analysis of Protein Expression in HEY Cells
2
Protein Extraction and Western Blot Analysis
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Total proteins of the cells were extracted with 100 μL of ice-cold lysate buffer (Beyotime, Shanghai, China) containing 1% phenylmethanesulfonyl fluoride and centrifuged at 12,000 g for 10 min. Protein concentrations were determined with a BCA Protein Assay Kit (Beyotime, Shanghai, China). After boiling for 10 min, the extracted protein was loaded at 50 μg/per lane and fractionated on SDS-PAGE gel, electrophoretically transferred to a polyvinylidene difluoride membrane (Millipore, Billerica, MA, USA). With the primary antibody incubation overnight and the secondary antibody incubation at 25 °C for 2 h, the bands were detected by super ELC detection reagent (Beyotime, Shanghai, China). Images were evaluated using Image-Pro Plus software (version 7.0, Olympus, Tokyo, Japan).
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