The IBRS-2 cells were placed at a 60 mm dish, then infected with SVA when they reached 80% confluence. Whole cellular lysates (30 μg protein) were subjected to immunoblot analysis with antibodies against STAT3, β-actin, phosphorylated-STAT3-Ser727, CDK2 (Abcam), and SVA-VP1 (self-made). Secondary antibodies were horseradish peroxidase-coupled anti-rabbit IgG antibodies (Thermo). Protein-bound antibodies were visualized with ECL plus detection reagents (Thermo). The protein expression was detected by ImageJ software and represented by area value.
Ecl plus detection reagents
Manufactured by Thermo Fisher Scientific
ECL plus detection reagents are chemiluminescent substrates used for the detection of proteins in Western blotting applications. The reagents produce a luminescent signal when exposed to the enzyme-labeled target protein, allowing for the visualization and quantification of the protein of interest.
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Lab products found in correlation
2 protocols using ecl plus detection reagents
1
SVA Infection and STAT3 Signaling
2
Lung Protein Extraction and Western Blot
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The total proteins were extracted from lungs using Protein Extraction Kit according to the manufacturer's protocol (Beyotime, Haimen, China). Protein concentrations were measured using the BCA protein assay kit. The protein samples with equal amounts (40 μg) of protein were separated on 12% SDS gel and transferred onto PVDF membranes. After blocking with 3% nonfat milk in TBST for 2 h at room temperature, the membrane was probed with the primary antibodies and HRP-conjugated secondary antibodies. The signals on the membranes were developed with the ECL Plus detection reagents (Thermo).
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