Total cell and tissue lysates were prepared in RIPA buffer (25 mM Tris-HCl, 150 mM NaCl, 1.0% NP-40, 1.0% deoxycholic acid, 0.1% SDS, 2 mM EDTA) containing 1mM PMSF, protease inhibitor cocktail and phosphatase inhibitor cocktail. Proteins were resolved by SDS-PAGE and immunoblotting was carried out using standard procedures. Membranes were incubated with primary antibodies against pAkt, Akt, pErk1/2, and Erk1/2(Cell Signaling, Danvers, MA), and α-tubulin (Active Motif, Carlsbad, CA) overnight at 4°C. HRP-conjugated secondary antibodies against rabbit and mouse IgG (GE Healthcare, Pittsburgh, PA) were used for protein detection. Densitometric analysis of immunoblots was performed using ImageJ software (v 1.47t, National Institutes of Health, Bethesda, MD).
Hrp conjugated secondary antibodies against rabbit and mouse igg
Manufactured by GE Healthcare
HRP-conjugated secondary antibodies against rabbit and mouse IgG are laboratory reagents used to detect and quantify primary antibodies targeting rabbit or mouse immunoglobulin G (IgG) in various immunoassays and immunochemistry techniques. These secondary antibodies are conjugated with the enzyme horseradish peroxidase (HRP), enabling signal amplification and detection.
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Lab products found in correlation
2 protocols using hrp conjugated secondary antibodies against rabbit and mouse igg
1
Immunoblotting Analysis of Cell Signaling
2
Immunoblotting Analysis of Cell Signaling
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Total cell and tissue lysates were prepared in RIPA buffer (25 mM Tris-HCl, 150 mM NaCl, 1.0% NP-40, 1.0% deoxycholic acid, 0.1% SDS, 2 mM EDTA) containing 1mM PMSF, protease inhibitor cocktail and phosphatase inhibitor cocktail. Proteins were resolved by SDS-PAGE and immunoblotting was carried out using standard procedures. Membranes were incubated with primary antibodies against pAkt, Akt, pErk1/2, and Erk1/2(Cell Signaling, Danvers, MA), and α-tubulin (Active Motif, Carlsbad, CA) overnight at 4°C. HRP-conjugated secondary antibodies against rabbit and mouse IgG (GE Healthcare, Pittsburgh, PA) were used for protein detection. Densitometric analysis of immunoblots was performed using ImageJ software (v 1.47t, National Institutes of Health, Bethesda, MD).
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