Western blotting was performed as previously described 56 (link), 61 (link). Briefly, cell lysates containing 50–100 μg of protein were resolved by electrophoresis on 10% SDS-polyacrylamide gels. Proteins were transferred to Hybond-P membranes, which were incubated in 5% non-fat dry milk for 1 hour and overnight at 4°C in 0.25% non-fat milk containing antibodies. The membranes were incubated for 1 hour at room temperature in 0.25% non-fat milk with an anti-rabbit or mouse IgG HRP-linked secondary antibody (Cell Signaling, Danvers, MA, USA). To ensure equal loading of proteins, the membranes were stripped under the same conditions as described above. They were then incubated with enhanced chemiluminescence reagents and exposed to X-ray film for 1–5 min.
Anti rabbit or mouse igg hrp linked secondary antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-rabbit or mouse IgG HRP-linked secondary antibody is a laboratory reagent used to detect and visualize target proteins in various immunoassays, such as Western blotting. The secondary antibody is conjugated with horseradish peroxidase (HRP), which enables the detection of the target protein through a colorimetric or chemiluminescent reaction.
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2 protocols using anti rabbit or mouse igg hrp linked secondary antibody
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Western Blotting Protocol for Protein Analysis
2
Western Blotting Protocol for Protein Analysis
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Western blotting was performed as previously described 56 (link), 61 (link). Briefly, cell lysates containing 50–100 μg of protein were resolved by electrophoresis on 10% SDS-polyacrylamide gels. Proteins were transferred to Hybond-P membranes, which were incubated in 5% non-fat dry milk for 1 hour and overnight at 4°C in 0.25% non-fat milk containing antibodies. The membranes were incubated for 1 hour at room temperature in 0.25% non-fat milk with an anti-rabbit or mouse IgG HRP-linked secondary antibody (Cell Signaling, Danvers, MA, USA). To ensure equal loading of proteins, the membranes were stripped under the same conditions as described above. They were then incubated with enhanced chemiluminescence reagents and exposed to X-ray film for 1–5 min.
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