Abstract
We prepared antibodies against phosphoserine (P-Ser) and phosphothreonine (P-Thr) by immunizing rabbits with P-Ser or P-Thr conjugated to bovine serum albumin. The antibodies (anti-P-Ser and anti-P-Thr) were purified using P-Ser or P-Thr affinity columns. Anti-P-Thr was highly specific for P-Thr, while anti-P-Ser showed weak cross-reactivity with P-Thr. We showed that these antibodies can immunodetect serine/threonine phosphorylated insulin and epidermal growth factor (EGF) receptors and several proteins which are phosphorylated on serine/threonine residues in response to insulin or EGF stimulation. The antibodies will certainly provide a good tool for discovering novel kinases and substrates involved in signal transduction.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies* / isolation & purification
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CHO Cells
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Chromatography, Affinity
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Cricetinae
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Epidermal Growth Factor / pharmacology*
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ErbB Receptors / metabolism*
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Humans
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Insulin / pharmacology*
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Kinetics
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Phosphates / metabolism
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Phosphoproteins / isolation & purification
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Phosphoproteins / metabolism*
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Phosphorylation
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Phosphoserine / analysis*
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Phosphoserine / immunology
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Phosphothreonine / analysis*
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Phosphothreonine / immunology
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Rabbits / immunology
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Receptor, Insulin / metabolism*
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Signal Transduction
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Tetradecanoylphorbol Acetate / pharmacology
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Transfection
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Tumor Cells, Cultured
Substances
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Antibodies
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Insulin
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Phosphates
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Phosphoproteins
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Phosphothreonine
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Phosphoserine
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Epidermal Growth Factor
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ErbB Receptors
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Receptor, Insulin
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Tetradecanoylphorbol Acetate