| 货号 | 9415S |
| 描述 | 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. |
| 反应种属 | Human/Mouse/Rat/All |
| 来源宿主 | Mouse |
| 应用 | IF-IC/F |
| 目标/特异性 | 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 Nos.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.) |
| 使用方法 | F(1:50) IF-IC (1:50) |
| 供应商 | CST |
| 标记 | Alexa Fluor 647 |
| 背景 | 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. |
| 存放说明 | 4C |
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。 |
