Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

ALK Activation Antibody Sampler Kit #8336

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-ALK (Tyr1586) (3B4) Rabbit mAb #3348 40 µl W H 80 (NPM-ALK) 220 (ALK) Rabbit IgG
Phospho-Jak3 (Tyr980/981) (D44E3) Rabbit mAb #5031 40 µl W H M (Mk) 115 Rabbit IgG
Phospho-Jak2 (Tyr1007) (D15E2) Rabbit mAb #4406 40 µl W H M (R) (Mk) (B) 125 Rabbit IgG
Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 40 µl W IP IHC-P IHC-F IF-IC F ChIP H M R Mk (Hm) (B) (Pg) (Hr) 79, 86 Rabbit IgG
Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb #4322 40 µl W IF-IC F H M (R) (Mk) (B) 90 Rabbit IgG
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 40 µl W IP IHC-P IF-IC F H M R Hm Mk Mi Dm Z B Dg Pg Sc (C) (Ce) 44, 42 Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 40 µl W IP IHC-P IHC-F IF-IC F H M R Hm Mk Dm Z B (C) (X) (Dg) (Pg) 60 Rabbit IgG
Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943 40 µl W IP H M R 60 Rabbit IgG
Phospho-PLCγ1 (Tyr783) Antibody #2821 40 µl W F H M R 155 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Mi=Mink  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  B=Bovine  Dg=Dog  Pg=Pig  Sc=S. cerevisiae  Hr=Horse  Ce=C. elegans
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

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.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF for the indicated times, using Phospho-PLCγ1 (Tyr783) Antibody #2821 (upper) or PLCγ1 Antibody #2822 (lower).

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).


Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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.


Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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.


Description

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.

Source / Purification

Activation state polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr783 of human PLCγ1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Rabbit monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr1586 of human ALK, Tyr1007 of human Jak2, Tyr980/981 of human and mouse Jak3, Tyr705 of mouse Stat3, Tyr694 of human Stat5a, Thr202/Tyr204 of human p44 MAP kinase, Ser473 of human Akt, or Tyr416 of human Src.

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).

  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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

* Product-specific protocol.

Companion Products

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

Products