Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Threonine (42H4) Mouse mAb #9386

general   kinase assay   motif   PBS   substrate   threonine  

No. Size Price
9386S 100 µl ( 10 western blots ) ¥2,846.00 现货查询 购买询价
9386 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 All Species Expected, Endogenous Mouse IgM
IP 1:50
E-P 1:2000

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, E-P=Peptide ELISA (DELFIA),

Specificity / Sensitivity

Phospho-Threonine (42H4) Mouse mAb binds phosphorylated threonine residues in a manner largely independent of the surrounding amino acid sequence. The antibody is phospho-specific but does not cross-react with phospho-tyrosine-containing sequences. It does show slight cross-reactivity with a few phospho-serine-containing peptides. By ELISA, it recognizes a wide variety of threonine-phosphorylated peptides. (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.)

Phospho-Threonine (42H4) Mouse mAb可以识别磷酸化苏氨酸残基,并且与周围氨基酸序列无关。该抗体是磷酸化特异性的,不与包含磷酸化酪氨酸的序列发生交叉反应。它与一些包含磷酸化丝氨酸的肽段有轻微交叉反应。在ELISA中,其可以识别多种苏氨酸磷酸化的肽段。(美国专利号:6,441,140、6,982,318、7,259,022、7,344,714;U.S.S.N. 11,484,485;及其对应的国外部分)

Source / Purification

Monoclonal antibody is produced by immunizing animals with phospho-Thr-containing peptides.

该单克隆抗体用包含磷酸化苏氨酸的肽段免疫动物制备。

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or treated with calyculin A, a threonine phosphatase inhibitor, or extracts from NIH/3T3 cells, untreated or treated with pervanadate, a tyrosine phosphatase inhibitor, using Phospho-Threonine (42H4) Mouse mAb (upper) or Phospho-Tyrosine Monoclonal Antibody (P-Tyr-100) #9411 (lower).

对A431细胞抽提液,未处理(-)或calyculin A(苏氨酸磷酸酶抑制剂)处理,或者对NIH/3T3细胞抽提液,未处理或过钒酸盐(酪氨酸磷酸酶抑制剂)处理,使用Phospho-Threonine (42H4) Mouse mAb (上图)或Phospho-Tyrosine Monoclonal Antibody (P-Tyr-100) #9411 (下图)进行Western blot分析。

ELISA-Peptide

ELISA-Peptide

Phospho-Threonine (42H4) Mouse mAb ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* denotes phosphorylated threonine.)

Phospho-Threonine (42H4) Mouse mAb ELISA分析:磷酸化和非磷酸化的信噪比。(T*代表磷酸化的苏氨酸)

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.

细胞内很多蛋白质的动态行为,包括分子相互作用的调节(1),亚细胞定位(2)和转录调节(3)都由多种转录后修饰调控(4)。对这些转录后修饰特异性的抗体对理解正常和病理的分子和细胞行为非常有价值。大部分蛋白修饰抗体设计出来与修饰的氨基酸残基发生反应(如磷酸化苏氨酸、磷酸化酪氨酸、乙酰化赖氨酸、硝基化酪氨酸),而不考虑这些残疾在序列中的位置。这种不考虑上下文而仅识别修饰后残基的能力给予这些抗体广泛的识别可能,赋予其识别上百种不同蛋白质的能力。这种广泛的识别使这些抗体在复杂分析和目标筛查的过程中具有价值。

Protein kinases are among the most abundant eukaryotic regulatory proteins; over 500 separate kinase genes are encoded in mammalian genomes (5,6). In spite of the importance of kinases in eukaryotic biology, relatively few of their physiological targets are known. Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381 and Phospho-Threonine (42H4) Monoclonal Antibody #9386 provide powerful tools for discovering targets of serine/threonine kinases, for monitoring and characterizing in vitro threonine phosphorylation reactions as well as for high throughput Ser/Thr kinase drug discovery.

蛋白激酶在是众多真核细胞调节蛋白之一;哺乳动物中有超过500个独立的激酶基因编码这些蛋白(5,6)。尽管它们在真核生物激酶中很重要,但对它们的生理靶标还相对知之甚少。Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381和Phospho-Threonine (42H4) Monoclonal Antibody #9386为发现丝氨酸/苏氨酸激酶、检测和鉴定体外苏氨酸磷酸化反应、高通量丝氨酸/苏氨酸激酶药物发现提供了有力工具。

  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.
  5. Venter, J.C. et al. (2001) Science 291, 1304-51.
  6. Manning, G. et al. (2002) Science 298, 1912-34.

Application References

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

Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 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 commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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