Phospho-Threonine Antibody (P-Thr-Polyclonal) (Magnetic Bead Conjugate) detects proteins and peptides phosphorylated at threonine residues in a manner largely independent of the surrounding amino acid sequence. The antibody is phospho-specific and may cross-react with some phospho-serine-containing sequences. By ELISA, it recognizes a wide variety of threonine-phosphorylated peptides, and by 2D gel Western blot analysis, it recognizes a large number of presumably threonine-phosphorylated proteins. Use of Phospho-Threonine-Proline Mouse mAb (P-Thr-Pro-101) #9391 is recommended to detect proteins containing threonine followed by proline.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic phospho-Thr-containing peptides. Antibodies are purified by protein A and peptide affinity chromatography.
Description
This Cell Signaling Technology antibody is immobilized by the covalent reaction of formylbenzamide-modified antibody with hydrazide-activated magnetic bead. Phospho-Threonine Antibody (P-Thr-Polyclonal) (Magnetic Bead Conjugate) is useful for immunoprecipitation assays. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381.
原厂资料：
Specificity / Sensitivity
Phospho-Threonine Antibody (P-Thr-Polyclonal) (Magnetic Bead Conjugate) detects proteins and peptides phosphorylated at threonine residues in a manner largely independent of the surrounding amino acid sequence. The antibody is phospho-specific and may cross-react with some phospho-serine-containing sequences. By ELISA, it recognizes a wide variety of threonine-phosphorylated peptides, and by 2D gel Western blot analysis, it recognizes a large number of presumably threonine-phosphorylated proteins. Use of Phospho-Threonine-Proline Mouse mAb (P-Thr-Pro-101) #9391 is recommended to detect proteins containing threonine followed by proline.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic phospho-Thr-containing peptides. Antibodies are purified by protein A and peptide affinity chromatography.
Description
This Cell Signaling Technology antibody is immobilized by the covalent reaction of formylbenzamide-modified antibody with hydrazide-activated magnetic bead. Phospho-Threonine Antibody (P-Thr-Polyclonal) (Magnetic Bead Conjugate) is useful for immunoprecipitation assays. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381.
Directions for Use
Add 10 μl of well-vortexed beads to 200 μl of cell lysate at 1 mg/ml in 1X Cell Lysis Buffer (10X) #9803. See protocol for more details.
For bead washing and subsequent elution of immunocomplexes, the beads can be separated from solution using our 6-Tube Magnetic Separation Rack #7017. Place the tubes containing the beads in the Magnetic Separation Rack and wait 1 to 2 minutes for the solution to clear before carefully removing the supernatant. Remove the tubes from the Magnetic Separation Rack, add new solution and resuspend the beads by gently vortexing or rocking the tube.
Background
Much of the dynamic behavior of cellular proteins, including the regulation of molecular interactions (1), subcellular localization (2), and transcriptional regulation (3) is controlled by a variety of post-translational modifications (4). Antibodies specific for these post-translational modifications are invaluable tools in the quest to understand normal and pathogenic molecular and cellular behavior.
General protein modification antibodies are designed to react with modified amino acid residues (e.g. phospho-threonine, phospho-tyrosine, acetyl-lysine, nitro-tyrosine) independently of the sequence in which they are embedded. This ability to recognize modified residues in a "context-independent" fashion gives these antibodies broad reactivities, presumably conferring upon them the ability to react with hundreds of distinct proteins. This broad pattern of reactivity makes these antibodies especially valuable in multiplex analyses and target discovery programs.
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