Cells were lysed in boiling Laemmli buffer supplemented with protease inhibitors, then sonicated and complemented with DTT. Protein concentration was determined by BCA (Thermo Scientific) assay. Fifteen to twenty micrograms of total protein were loaded onto SDS-PAGE gels and transferred onto nitrocellulose membranes. The membrane was blocked with TBST (1× TBS with 0.1% Tween 20) + 5% milk at room temperature for 1 h and incubated with primary antibody and then with horseradish peroxidase (HRP)-coupled secondary antibody (Amersham, Piscataway, NJ, United States). Activity was visualized by electrochemiluminescence. Antibodies used in this study are anti-Histone H3 (Cell signaling Technology #9717) and anti-Acetyl-Histone H3 (Lys9) (Cell signaling Technology #9649).
Horseradish peroxidase hrp coupled secondary antibody
Manufactured by Cytiva
Sourced in United States
Horseradish peroxidase (HRP)-coupled secondary antibody is a laboratory reagent used in various immunoassay techniques. It consists of a secondary antibody that is conjugated with the enzyme horseradish peroxidase. This conjugate can be used to detect and quantify target proteins in samples by catalyzing a colorimetric or chemiluminescent reaction.
Automatically generated - may contain errors
2 protocols using horseradish peroxidase hrp coupled secondary antibody
1
Histone H3 Acetylation Immunoblotting
2
Histone H3 Acetylation Immunoblotting
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