Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Threonine Antibody (P-Thr-Polyclonal) (HRP Conjugate) #6949

Applications Reactivity Sensitivity Source
W E-P Endogenous Rabbit

Applications Key:  W=Western Blotting  E-P=ELISA (Peptide)
Reactivity Key:
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

* Product-specific protocol.

Specificity / Sensitivity

Phospho-Threonine Antibody (P-Thr-Polyclonal) (HRP 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. We recommend the use of Phospho-Threonine-Proline Mouse mAb (P-Thr-Pro-101) #9391 to detect proteins containing threonine followed by proline. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with Calyculin A #9902 (0.1 µM for 45 minutes), using Phospho-Threonine Antibody (P-Thr-Polyclonal) (HRP Conjugate).

ELISA-Peptide

ELISA-Peptide

Signal-to-noise (S/N) values of phosphopeptide versus nonphosphopeptide signals using LumiGLO® and Peroxide Reagents #7003.

Description

This Cell Signaling Technology® antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381.

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.

  1. Yaffe, M.B. and Elia, A.E. (2001) Curr Opin Cell Biol 13, 131-8.
  2. Appella, E. and Anderson, C.W. (2001) Eur J Biochem 268, 2764-72.
  3. Jenuwein, T. and Allis, C.D. (2001) Science 293, 1074-80.
  4. Krishna, R.G. and Wold, F. (1993) Adv Enzymol Relat Areas Mol Biol 67, 265-98.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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