Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) #9415

motif   phosphotyrosine   PY100   substrate   tyrosine  

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse,Rat,All Species Expected, Endogenous Mouse IgG1
IF-IC 1:50

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Phospho-Tyrosine Mouse mAb (P-Tyr-100) is a high affinity antibody. ELISAs against a wide variety of phosphopeptides indicate that (a) P-Tyr-100 binds phospho-Tyr in a manner largely independent of the surrounding amino acid sequence. 2D gel Western blot analysis of pervanadate-treated cell extracts also shows that P-Tyr-100 interacts with a broad range of tyrosine-phosphorylated proteins. P-Tyr-100 does not cross-react with peptides containing phospho-Ser or phospho-Thr. (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-Tyrosine Mouse mAb (P-Tyr-100)磷酸化酪氨酸鼠单抗亲和力非常高。ELISA中对大量磷酸化肽段的实验显示P-Tyr-100结合磷酸化酪氨酸,不受周围氨基酸序列影响。二维凝胶Western Blot分析过钒酸盐(pervanadate)处理的细胞抽提物也显示P-Tyr-100与大量酪氨酸磷酸化的蛋白质发生相互作用。P-Tyr-100不与其它含磷酸化丝氨酸或磷酸化苏氨酸的肽段发生相互作用。(美国专利号:6,441,140、6,982,318、7,259,022、7,344,714;U.S.S.N. 11,484,485;及所有国外相应专利)

Source / Purification

Monoclonal antibody is produced by immunizing animals with phospho-tyrosine-containing peptides . The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6.

该单克隆抗体用含磷酸化酪氨酸的肽段免疫动物制备。抗体在最优条件下以F/P比2-6被偶联到Alexa Fluor® 647。


This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411.

CST公司生产的抗体在最优条件下与Alexa Fluor® 647 荧光染料结合,已通过在人类细胞上进行直接流式细胞仪分析的内部测试。抗体预计可以和未结合的Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411一样具有相同的交叉反应活性。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K562 cells, untreated (green) or Gleevec®- treated (blue), using Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluor® 647 Conjugate) compared with a nonspecific negative control antibody (red).

对K562细胞,未处理(绿)或Gleevec®处理(蓝),使用Phospho-Tyrosine Mouse mAb (P-Tyr-100) (Alexa Fluo® 647 Conjugate)和非特异性阴性对照抗体(红色),进行流式细胞仪分析。



Confocal immunofluorescent image of two Gefinitib (Iressa) treated non-small cell lung cancer cell lines. HCC827 cells have the E746_A750 deletion in exon 19 of the EGFR gene, and are highly sensitive to Gefitinib. H1975 cells have the (T790M) mutation that confers Gefitinib-resistance. Both cell lines were treated and then double-labeled with Phospho-S6 Ribosomal Protein Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854 and Phospho-Tyrosine Mouse mAb (Alexa Fluor® 647 Conjugate) #9415. Untreated HCC827 (A) and H1975 (C) cells show bright phospho-S6 (green) and phospho-tyrosine (red pseudocolor) label. Phospho-S6 and phospho-tyrosine signals dramatically decrease following Gefitinib treatment in HCC827 cells (B), with little or no change in the Gefitinib-resistant H1975 cells (D).

两个Gefinitib (Iressa)处理的非小细胞肺癌细胞系的共聚焦免疫荧光照片。HCC827细胞在EGFR基因19号外显子有E746_A750缺失,对Gefitinib高度敏感。H1975细胞有(T790M)突变则会对Gefitinib抵抗。两个细胞系都被处理过,然后用Phospho-S6 Ribosomal Protein Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854和Phospho-Tyrosine Mouse mAb (Alexa Fluor® 647 Conjugate) #9415进行双重标记。未处理HCC827(A)和H1975(C)细胞显示出明亮的phospho-S6(绿)和phospho-tyrosine(伪彩红)标签。HCC827细胞(B)的Phospho-S6和phospho-tyrosine信号在Gefitinib处理后显著下降,在Gefitinib抵抗的H1975细胞(D)则几乎没有变化。


Tyrosine phosphorylation plays a key role in cellular signaling (1). In cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (2). Antibodies specific for phospho-tyrosine (3,4) have been invaluable reagents in these studies. The phospho-tyrosine monoclonal antibodies developed by Cell Signaling Technology provide exceptionally sensitive new tools of increased utility for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery.

酪氨酸磷酸化在细胞信号转导中扮演重要角色(1)。在肿瘤中,失调的酪氨酸激酶活性可以通过产生不适当的增生和存活信号而导致恶性肿瘤和肿瘤形成(2)。具有磷酸化酪氨酸特异性的抗体在这些研究中很有价值(3,4)。Cell Signaling Technology开发的磷酸化酪氨酸单克隆抗体在高通量药物发现中为研究酪氨酸磷酸化和监测酪氨酸激酶活性提供了卓越的高灵敏的新工具。

  1. Schlessinger, J. (2000) Cell 103, 211-25.
  2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.
  3. Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9.
  4. Glenney, J.R. et al. (1988) J Immunol Methods 109, 277-85.

Application References

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Companion Products

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

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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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