Myocardial tissue samples were homogenized with lysis buffer, and the protein was extracted. Protein concentrations were measured by the bicinchoninic acid method. The samples were separated by SDS-PAGE and identified with following antibodies: PI3K, phosphorylated PI3K (p-PI3K), Akt and p-Akt (Santa Cruz Biotechnology). After incubation with HRP-coupled secondary antibody (Santa Cruz Biotechnology), the bands were visualized with a chemiluminescence developing agents (Amersham Biosciences, UK). GAPDH was probed in the blots as internal control for loading. Results were semi-quantified as optical density band area by the Image J.
Chemiluminescence developing agents
Manufactured by Cytiva
Sourced in United Kingdom
Chemiluminescence developing agents are chemical compounds used in various laboratory applications to detect and quantify specific biomolecules or cellular processes. These agents emit light upon a chemical reaction, allowing for the visualization and analysis of target analytes.
Automatically generated - may contain errors
Lab products found in correlation
P akt, by Santa Cruz Biotechnology (1 mentions)
Hrp coupled secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase coupled secondary antibody, by Santa Cruz Biotechnology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Cst2532, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Sodium dodecyl sulfate polyacrylamide gel electrophoresis, by Bio-Rad (1 mentions)
3 protocols using chemiluminescence developing agents
1
Myocardial Protein Expression Analysis
2
Western Blot Protein Analysis Protocol
3
Quantifying AMPK Activation in Liver Tissue
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The total protein concentration from liver tissues were measured by the bicinchoninic acid method. To determine the protein levels of AMPKα and phospho-AMPKα, equal amounts of protein extract (50 µg) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Bio-Rad, CA, USA) and transferred electrophoretically to nitrocellulose membranes. The membranes were then blocked with 5% bovine serum albumin in Tris-buffered saline Tween (20 mM Tris, (pH 7.6), 137 mM NaCl, and 0.1% Tween 20). The primary antibodies against AMPKα and phospho-AMPKα were obtained from Cell Signalling Technology, Inc. (Beverly, MA, USA) with the catalog number #CST-2532 and #CST-2535, respectively. All of the antibodies were used at a dilution of 1:1000. The membrane was incubated overnight at 4 °C with the primary antibody, and the bound antibody was visualized using the respective horseradish peroxidase-conjugated secondary antibodies (Cell Signalling Technology, Inc. (Beverly, MA, USA) catalog #CST-5127)) and chemiluminescence developing agents (Amersham Biosciences, Buckinghamshire, UK).
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