Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb #14678

No. Size Price
14678S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
14678 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 80 (NPM-ALK), 220 (ALK) Rabbit IgG
IP 1:100
F 1:200
IF-IC 1:400

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat,

Specificity / Sensitivity

Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb recognizes endogenous levels of ALK protein only when phosphorylated at Tyr1507 (equivalent to Tyr567 of NPM-ALK).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1507 of human ALK protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of KARPAS-299 cells, untreated (left) or treated with Crizotinib #4401 (1 μM, 2hr; center), and HeLa cells (right), using Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of KARPAS-299 cells, untreated (green) or treated with Crizotinib #4401 (1 μM, 2 hr, 37ºC; blue), using Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Western Blotting

Western Blotting

Western blot analysis of extracts from KARPAS-299 cells, untreated (-) or treated with Crizotinib #4401 (1 μM, indicated times), using Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb (upper) and ALK (C26G7) Rabbit mAb #3333 (lower). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

IP

IP

Immunoprecipitation of phospho-ALK from KARPAS-299 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb. Western blot analysis was performed using Phospho-ALK (Tyr1507) (D6F1V) Rabbit mAb. KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Background

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 a nucleophosmin (NPM)-ALK fusion protein produced by a translocation (4). Investigators have found that the NPM-ALK fusion protein is a constitutively active, oncogenic tyrosine kinase associated with anaplastic lymphoma (4). Research literature suggests that activation of PLCγ by NPM-ALK may be a crucial step for its mitogenic activity and involved in the pathogenesis of anaplastic lymphomas (5).

A distinct ALK oncogenic fusion protein involving ALK and echinoderm microtubule-associated protein like 4 (EML4) has been described in the research literature from a non-small cell lung cancer (NSCLC) 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 (6-8).

Phosphorylation of ALK on Tyr1507 was identified at Cell Signaling Technology using PhosphoScan®, an LC-MS/MS platform used for phosphorylation site discovery (6). Phosphorylation of ALK at Tyr1507 (Tyr567 in NPM-ALK) has been shown to be important for interaction with the adaptor proteins Shc, FRS2-α, and FRS2-β (9,10).

  1. Stoica, G.E. et al. (2001) J Biol Chem 276, 16772-9.
  2. Iwahara, T. et al. (1997) Oncogene 14, 439-49.
  3. Morris, S.W. et al. (1997) Oncogene 14, 2175-88.
  4. Morris, S.W. et al. (1994) Science 263, 1281-4.
  5. Bai, R.Y. et al. (1998) Mol Cell Biol 18, 6951-61.
  6. Rikova, K. et al. (2007) Cell 131, 1190-203.
  7. Takeuchi, K. et al. (2008) Clin Cancer Res 14, 6618-24.
  8. Soda, M. et al. (2007) Nature 448, 561-6.
  9. Turner, S.D. et al. (2007) Cell Signal 19, 740-7.
  10. Chikamori, M. et al. (2007) Oncogene 26, 2950-4.

Application References

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

DRAQ5 is a registered trademark of Biostatus Limited.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

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

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

PhosphoScan is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Tween is a registered trademark of ICI Americas, Inc.

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

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