CST Guide:

Section I: Research Areas
chapter 02: Signaling
ALK, an RTK that forms oncogenic fusion proteins with
EML4, TFG, and KIF5B, is expressed in lung carcinoma.
ALK (D5F3 ® ) XP ® Rabbit mAb #3633: IHC
analysis of paraffin-embedded human lung
carcinoma with high (left) and low (right) levels
of ALK expression using #3633.
Phospho-Jak2 (Tyr1008) (D4A8) Rabbit
mAb #8082: WB analysis of extracts from
BaF3 cells (A), untreated or treated with Mouse
Interleukin-3 (mIL-3) #8923 (10 ng/ml, 5 min),
using #8082 (upper) or total Jak2 (D2E12) XP ®
Rabbit mAb #3230 (lower).
Jak2 (D2E12) XP ® Rabbit mAb #3230:
Confocal IF analysis of K-562 cells (B) using
#3230 (green). Blue pseudocolor = DRAQ5 ®
(fluorescent DNA dye).
A
kDa
200
140
100
80
60
50
40
30
200
140
100
Phospho-
Jak2
(Tyr1008)
Jak2
B
Treatment with
IL-3 results in
phosphorylation
of Jak2 at Tyr1008.
80
60
50
40
Ligand activation of
Met, a high affinity RTK
for hepatocyte growth
factor (HGF), results
in phosphorylation at
Tyr1234/1235.
A kDa
200
140
100
80
60
50
40
Phospho-Met
(Tyr1234/5)
B
Phospho-Met (Tyr1234/1235) (D26) XP ® Rabbit mAb #3077: WB analysis of cell
extracts from HeLa cells, untreated or stimulated with HGF (A), using #3077 (upper) and Met
(25H2) Mouse mAb #3127 (lower). Confocal IF analysis of MKN-45 cells, untreated (B) or
treated with SU11274 (1 μM, 3 hr) (C), using #3077. Blue pseudocolor = DRAQ5 ® #4084
(fluorescent DNA dye).
C
Treatment with a
Met inhibitor prevents
Met activation and
phosphorylation at
Tyr1234/1235.
The nonreceptor tyrosine kinase
Zap-70 is expressed in T cells.
30
– +
Zap-70 (D1C10E) XP ® Rabbit mAb (Alexa Fluor ® 488 Conjugate) #9473:
Flow cytometric analysis of Ramos (B cells; blue) and Jurkat (T cells; green) cells
using #9473.
mIL-3
Events
Zap-70
200
140
100
80
60
50
40
– +
Met
HGF
Nonreceptor Tyrosine Kinases
The cytoplasmic nonreceptor tyrosine kinases transduce signals generated by RTKs and other receptors
from the cell surface to the nucleus. Receptor-associated tyrosine kinases are phosphorylated within the
activation loop of their kinase catalytic domain, resulting in activation and propagation of downstream
signals through phosphorylation of other kinases, adaptor proteins, or transcription factors. In addition
to their kinase domain, nonreceptor tyrosine kinases often contain various domains that mediate
protein-protein or protein-lipid interactions, which allows association with activated receptors or downstream
signaling nodes. For example, SH2 domains bind phospho-tyrosine residues, while PH domains
direct interactions with membrane lipids.
PDGF treatment results in phosphorylation
of Src family proteins at Tyr416.
Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb #6943: WB 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 #6943 (upper) or Src (36D10) Rabbit mAb
#2109 (lower).
Select Reviews
Arteaga, C.L. and Engelman, J.A. (2014) Cancer Cell 25, 282−303. • Davies, K.D. and Doebele, R.C. (2013) Clin. Cancer
Res. 19, 4040−4045. • Hallberg, B. and Palmer, R.H. (2013) Nat. Rev. Cancer 13, 685−700. • Liu, S.T., Pham, H., Pandol,
S.J., and Ptasznik, A. (2014) Front. Physiol. 4, 416. • Miaczynska, M. (2013) Cold Spring Harb. Perspect. Biol. 5, a009035.
• Panjarian, S., Lacob, R.E., Chen, S., Engen, J.R., and Smithgall, T.E. (2013) J. Biol. Chem. 288, 5443−5450. • Shaw, A.T.,
Hsu, P.P., Awad, M.M., and Engelman, J.A. (2013) Nat. Rev. Cancer 13, 772−787.
kDa
100
80
60
50
40
30
20
100
80
60
50
40
30
20
– +
Phospho-Src
(Tyr416)
Src
hPDGF-BB
Commonly Studied Tyrosine Kinases Targets
Target M P E S C
ACPP
•
ALK • • •
Phospho-ALK (Tyr1078) • •
Phospho-ALK (Tyr1096) • •
Phospho-ALK (Tyr1278) • •
Phospho-ALK (Tyr1282/Tyr1283) • •
•
Phospho-ALK (Tyr1278/Tyr1282/
Tyr1283)
Phospho-ALK (Tyr1586) • • •
Phospho-ALK (Tyr1604) • • •
Axl
• • • •
Phospho-Axl (pan Tyr)
•
Phospho-Axl (Tyr702) •
DDR1
• • •
Phospho-DDR1 (pan Tyr)
•
Phospho-DDR1 (Tyr792) •
DDR2
•
EGFR
• • • • •
Phospho-EGFR (Thr669) • •
Phospho-EGFR (Tyr845) • • •
Phospho-EGFR (Tyr992) •
Phospho-EGFR (Tyr998) •
Phospho-EGFR (Tyr1045) •
Phospho-EGFR (Ser1046/1047) •
Phospho-EGFR (Tyr1068) • • • •
Phospho-EGFR (Tyr1086) •
Phospho-EGFR (Tyr1148) •
Phospho-EGFR (Tyr1173) • •
• • •
EGF Receptor
(E746-A750del Specific)
Target M P E S C
EGF Receptor (L858R Mutant Specific) • •
EphA2
•
Phospho-EphA2 (Tyr588) •
Phospho-EphA2 (Tyr594) •
Phospho-EphA2 (Tyr772) •
Phospho-EphA2 (Ser897) •
Phospho-EphA3 (Tyr779) •
EphA3/EphA4/EphA5 •
EphB1
•
EphB6
•
Phospho-Ephrin B (Tyr324/329) •
Eps8
•
EREG
•
Etk/BMX
•
Phospho-Etk/BMX (Tyr40) •
FGFR1 • • • •
FGFR1 (pan Tyr)
•
FGFR1 (Tyr653/Tyr654) • • •
FGFR1 (Tyr766)
•
FGFR2 • • •
Phospho-FGFR2 (Pan Tyr)
•
FGFR3
•
FGFR4 • • • •
FLT3 • •
Phospho-FLT3 (pan Tyr)
•
Phospho-FLT3 (Tyr589/Tyr591) •
Phospho-FLT3 (Tyr591) • • •
Phospho-FLT3 (Tyr842) •
Phospho-FLT3 (Tyr969) •
HER2/ErbB2
• • • • •
These protein targets represent key
nodes within tyrosine kinases signaling
pathways and are commonly studied
in tyrosine kinases research. Primary
antibodies, antibody conjugates, and
antibody sampler kits containing these
targets are available from CST.
Listing as of September 2014. See our
website for current product information.
M Monoclonal Antibody
P Polyclonal Antibody
E PathScan ® ELISA Kits
S SignalSilence ® siRNA
C Antibody Conjugate
68 For Research Use Only. Not For Use in Diagnostic Procedures. See pages 302 & 303 for Pathway Diagrams, Application, and Reactivity keys.
www.cellsignal.com/csttyrosine 69