| 货号 | 8336T |
| 描述 | The ALK Activation Antibody Sampler Kit provides an economical means to evaluate the activation status of multiple members of the ALK pathway, including phosphorylated ALK, Jak2, Jak3, Stat3, Stat5, PLCγ1, Akt, Src, and p44/42 MAPK. The kit includes enough antibody to perform four western blot experiments with each primary antibody. |
| 目标/特异性 | Each antibody in the ALK Activation Antibody Sampler Kit recognizes the phosphorylated form of its specific target. Phospho-ALK (Tyr1586) (3B4) Rabbit mAb may cross-react with other activated protein tyrosine kinases including EGFR. Phospho-Jak2 (Tyr1007) (D15E2) Rabbit mAb may cross react with phosphorylated Jak3 and Tyk2. Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb may cross react with overexpressed phosphorylated RTKs. |
| 供应商 | CST |
| 背景 | Anaplastic lymphoma kinase (ALK) is a tyrosine kinase receptor for pleiotrophin (PTN), a growth factor involved in embryonic brain development (1-3). In ALK-expressing cells, PTN induces phosphorylation of both ALK and the downstream effectors IRS-1, Shc, PLCγ, and PI3 kinase (1). ALK was originally discovered as an NPM (nucleophosmin)-ALK fusion protein produced by a translocation (4). The NPM-ALK fusion protein is a constitutively active, oncogenic tyrosine kinase associated with anaplastic lymphoma (4). Activation of PLCγ by NPM-ALK has been suggested to be a crucial step for its mitogenic activity and may be important in the pathogenesis of anaplastic lymphomas (5). A distinct ALK oncogenic fusion protein involving ALK and EML4 has been described from a non-small cell lung cancer cell line, with corresponding fusion transcripts present in some cases of lung adenocarcinoma. The short, amino-terminal region of the microtubule-associated protein EML4 is fused to the kinase domain of ALK (6,7). |
| 存放说明 | -20C |
| 参考文献 | Stoica, G.E. et al. (2001) J Biol Chem 276, 16772-9. Iwahara, T. et al. (1997) Oncogene 14, 439-49. Morris, S.W. et al. (1997) Oncogene 14, 2175-88. Morris, S.W. et al. (1994) Science 263, 1281-4. Bai, R.Y. et al. (1998) Mol Cell Biol 18, 6951-61. Rikova, K. et al. (2007) Cell 131, 1190-203. Takeuchi, K. et al. (2008) Clin Cancer Res 14, 6618-24. Soda, M. et al. (2007) Nature 448, 561-6. |
Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or stimulated with hPDGF-BB #8912 (5 min; +), and from A-431 cells, untreated (-) or stimulated with hEGF #8916 (5 min; +), using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Immunoprecipitation of phospho-PLCγ1 (Tyr783) from A-431 cell extracts stimulated with hEGF #8916 using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb. | |
Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or treated with recombinant mouse PDGF-BB (200 ng/ml, 15 min; green), using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Confocal immunofluorescent analysis of A-431 cells, untreated (left), treated with hEGF #8916 (100 ng/ml, 5 min; center), or treated with hEGF #8916 (100 ng/ml, 5 min) and λ phosphatase (right), using Phospho-PLCγ1 (Tyr783) (D6M9S) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Immunohistochemical analysis using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb on SignalSlide® Phospho-Stat1/3/5 IHC Controls #8105 (paraffin-embedded HeLa cell pellets, untreated (left), treated with Human Interferon-α1 (hIFN-α1) #8927 (middle), or treated with Human Epidermal Growth Factor (hEGF) #8916 (right). | |
Western blot analysis of extracts from UT-7 cells untreated (-) or treated (+) with erythropoietin (EPO, 3 units/ml, 5 min), TF-1 cells untreated (-) or treated (+) with Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF) #8922 (100 ng/ml, 10 min), and NK-92 cells untreated (-) or treated (+) with Human Interleukin-2 (hIL-2) #8907 (100 ng/ml, 10 min), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb #4322 (upper) or total Stat5 (3H7) Rabbit mAb #9358 (lower).用Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb #4322 (上) 或total Stat5 (3H7) Rabbit mAb #9358 (下) 对以下细胞的提取物进行免疫印迹检测： 未处理的（-)或经过erythropoietin (EPO, 3 units/ml, 5 min)处理的（+）UT-7细胞，未处理的（-)或经过Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF) #8922 (100 ng/ml, 10 min)处理的（+）TF-1细胞，未处理的（-)或经过Human Interleukin-2 (hIL-2) #8907 (100 ng/ml, 10 min)处理的（+）NK-92细胞。 | |
Western blot analysis of extracts from NIH/3T3 cells, serum-starved (-) or treated (+) with Human Platelet-Derived Growth Factor BB (hPDGF-BB) #8912 (100 ng/ml, 15 min), using Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943 (upper) or Src (36D10) Rabbit mAb #2109 (lower). 用Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943 (上) 或Src (36D10) Rabbit mAb #2109 (下) 对以下细胞的提取物进行免疫印迹检测：血清饥饿处理的(-)或Human Platelet-Derived Growth Factor BB (hPDGF-BB) #8912 (100 ng/ml, 15 min)处理的(+)NIH/3T3 细胞。 | |
Western blot analysis of extracts from PC-3 cells, untreated (-) or LY294002/wortmannin-treated (+), and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).用Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (上) 或Akt (pan) (C67E7) Rabbit mAb #4691 (下) 对以下细胞的提取物进行免疫印迹检测： 未处理的（-)或经过LY294002/wortmannin处理的（+）PC-3细胞，血清饥饿处理的或PDGF处理的NIH/3T3 细胞。 | |
Western blot analysis of extracts from 293, NIH/3T3, and C6 cells treated with λ phosphatase or TPA as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).用phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (上) 或p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (下) 对以下细胞的提取物进行免疫印迹检测：经过λ phosphatase 或TPA 处理的293, NIH/3T3和 C6 细胞（详见注释）。 | |
Western blot analysis of extracts from UT-7 cells, untreated (-) or treated (+) with erythropoietin (EPO, 3 units/ml, 5 min) and from untreated HEL cells, using Phospho-Jak2 (Tyr1007) (D15E2) Rabbit mAb #4406 (upper) or total Jak2 (D2E12) XP® Rabbit mAb #3230 (lower). HEL cells contain a V617F activating mutation in Jak2.用Phospho-Jak2 (Tyr1007) (D15E2) Rabbit mAb #4406 (上) 或total Jak2 (D2E12) XP® Rabbit mAb #3230 (下) 对以下细胞的提取物进行免疫印迹检测：未处理的(-) 或erythropoietin (EPO, 3 units/ml, 5 min)处理的(+)UT-7细胞，未处理的HEL细胞。 | |
Western blot analysis of extracts from NK-92 cells, untreated (-) or treated (+) with Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, 15 min), using Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb #5031 (upper) or Jak3 Antibody #3775 (lower).用Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb #5031 (上) 或 Jak3 Antibody #3775 (下) 对未处理的或Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, 15 min)处理的NK-92细胞的提取物进行免疫印迹检测。 | |
Western blot analysis of extracts from Jurkat and HeLa cells untreated (-) or treated (+) with IFN-α (left), as well as A431 cells untreated (-) or EGF-treated (+) (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145. Note that basal phospho-Stat3 in A431 cells is detected by this antibody.用Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145对如下处理的细胞的裂解液进行免疫印迹检测：左，未经处理（-）或用IFN-α处理的（+）Jurkat 和 HeLa细胞；右，未经处理（-）或用EGF处理的（+）A431细胞。注意：本抗体可以检测到A431细胞中phospho-Stat3的本底表达。 | |
Western blot analysis of extracts NIH/3T3 cells, serum-starved or treated with human Platelet-Derived Growth Factor BB hPDGF-BB #8912 (100 ng/ml, 15 min), using Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb (upper) or Src (36D10) Rabbit mAb #2109 (lower). | |
Western blot analysis of SUP-M2 cell lysates untreated (-) or treated (+) with calf intestinal phosphatase (CIP), using Phospho-ALK (Tyr1586) (3B4) Rabbit mAb #3348 (A and B), and ALK Antibody (C and D).用Phospho-ALK (Tyr1586) (3B4) Rabbit mAb #3348 (A 和B)和ALK Antibody (C 和D)对如下处理的SUP-M2细胞的裂解液进行免疫印迹检测：未经处理（-），calf intestinal phosphatase (CIP)处理的（+）。 |
