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Immunoprecipitation Image 1: Phospho-Threonine Antibody (P-Thr-Polyclonal) (Sepharose® Bead Conjugate) — Immunoprecipitation of Jurkat cell extracts, untreated (-) or calyculin A-treated (+), using Phospho-Threonine Antibody (P-Thr-Polyclonal) (Sepharose^® Bead Conjugate). The western blot was probed using Phospho-Threonine Antibody (P-Thr-Polyclonal) (HRP Conjugate) #6949.

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Supporting Data

REACTIVITY	H All
SENSITIVITY	Endogenous
MW (kDa)
SOURCE	Rabbit

Application Key:

IP-Immunoprecipitation

Species Cross-Reactivity Key:

H-Human
All-All Species Expected

Product Information

Product Description

This Cell Signaling Technology antibody is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated Sepharose^® beads. Phospho-Threonine Antibody (P-Thr-Polyclonal) (Sepharose^® Bead Conjugate) is useful for immunoprecipitation assays. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381.

Product Usage Information

Application	Dilution
Immunoprecipitation	1:20

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol. Store at –20°C. Do not aliquot the antibodies.

Protocol

Specificity / Sensitivity

Phospho-Threonine Antibody (P-Thr-Polyclonal) (Sepharose^® Bead Conjugate) immunoprecipitates 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. CST recommends the use of Phospho-Threonine-Proline 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.)

Species Reactivity:

Human, All Species Expected

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.

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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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

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Phospho-Threonine Antibody (P-Thr-Polyclonal) (Sepharose® Bead Conjugate) #8781