Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Thr) MAPK/CDK Substrate Mouse mAb #2321

Applications Reactivity Source Isotype
W IHC-P E-P All Mouse IgM

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Phospho-(Thr) MAPK/CDK Substrate Mouse mAb detects phospho-threonine only when followed by proline. It reacts with proteins/peptides phosphorylated on the Thr-Pro motif in an otherwise highly context-independent fashion. The antibody does not cross-react with phospho-threonine in the absence of an adjacent proline. The antibody does not cross-react with phospho-tyrosine, but does react with some phospho-serine peptides containing the phospho-serine-proline motif (e.g., phospho-Elk-1). (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

Monoclonal antibody is produced by immunizing mice with synthetic phospho-threonine-proline-containing peptides (KLH-coupled). This antibody is a mouse IgM clone and can be recognized by anti-mouse Ig (whole molecule) secondary antibody. Antibody is purified by protein A chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or nocodazole-treated (1 µg/ml for 12 hours prior to lysis) and subjected to 2-D gel electrophoresis, using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or treated with serum and okadaic acid, using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb (left). The right panel shows total protein staining.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder, using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analyisis of paraffin-embedded human breast carcinoma, showing staining of proteins containing phospho-threonine-proline motifs, using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb.


ELISA-Peptide

ELISA-Peptide

Signal-to-noise ratio of ELISA reading using Phospho-(Thr) MAPK/CDK Substrate Mouse mAb with phospho-Thr-Pro- containing peptides (T*P) and non-Thr-Pro-containing peptides. (T* denotes phosphorylated threonine.)

Background

The MAPK and CDK families of serine/threonine protein kinases play important roles in cell signaling and cell cycle control. These kinases phosphorylate threonine or serine followed by a proline residue (1-6). To study and discover new MAPK and CDK substrates, CST has developed antibodies that bind to phospho-threonine followed by proline.

As determined by ELISA using a wide variety of phospho-Thr-Pro peptides, Phospho-(Thr) MAPK/CDK Substrate Monoclonal Antibody recognizes the phospho-Thr-Pro motif in a highly context-independent fashion. It also interacts with a broad range of phospho-Thr-Pro-containing proteins as determined by Western blot analysis of nocodazole-treated Jurkat cell extracts resolved on a 2-D gel.

  1. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  2. Seger, R. and Krebs, E.G. (1995) FASEB J 9, 726-35.
  3. Nurse, P. (2000) Cell 100, 71-8.
  4. Cross, T.G. et al. (2000) Exp Cell Res 256, 34-41.
  5. Yang, C.C. et al. (1998) J Protein Chem 17, 329-35.
  6. Reynolds, C.H. et al. (2000) J Neurochem 74, 1587-95.

Application References

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Companion Products

License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.

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