Download EphA2 (D4A2) XP® Rabbit mAb

Store at -20°C
EphA2 (D4A2) XP® Rabbit mAb
#6997
Orders n 877-616-CELL (2355)
[email protected]
Support n 877-678-TECH (8324)
[email protected]
Web n www.cellsignal.com
rev. 03/28/16
For Research Use Only. Not For Use In Diagnostic Procedures.
Entrez-Gene ID #1969
Swiss-Prot Acc. #P29317
Applications
W, IP, IHC-P, IF-IC
Endogenous
Species Cross-Reactivity*
H, M, R, Mk
Molecular Wt.
125 kDa
Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibody.
Isotype
Rabbit IgG**
*Species cross-reactivity is determined by western blot.
© 2014 Cell Signaling Technology, Inc.
XP® and Cell Signaling Technology® are trademarks of Cell Signaling Technology, Inc.
-3
S7
IM
CD
200
140
100
80
Recommended Antibody Dilutions:
Western blotting
1:1000
Immunoprecipitation1:100
Immunohistochemistry (Paraffin)
1:200†
Unmasking buffer: EDTA
Antibody diluent: SignalStain® Antibody Diluent #8112
Detection reagent: SignalStain® Boost (HRP, Rabbit) #8114
†Optimal IHC dilutions determined using SignalStain® Boost IHC
Detection Reagent.
Immunofluorescence (IF-IC)1:200
EphA2
60
50
40
30
20
For application specific protocols please see the web
page for this product at www.cellsignal.com.
10
Western blot analysis of extracts from various cell lines using
EphA2 (D4A2) XP® Rabbit mAb
Please visit www.cellsignal.com for a complete listing
of recommended companion products.
Background References:
(1) Wilkinson, D.G. (2000) Int. Rev. Cytol. 196, 177-244.
A549
(2) Klein, R. (2001) Curr. Opin. Cell Biol. 13, 196-203.
(3) Dodelet, V.C. and Pasquale, E.B. (2000) Oncogene 19,
5614-5619.
EphA2 is overexpressed in various tumor cells, and it has
been suggested that EphA2 may promote malignancy.
However, several studies demonstrate that EphA2 plays an
important role in tumor suppression (8). The role of EphA2
in tumor development may depend upon regulation of its
tyrosine kinase activity.
(4) Holder, N. and Klein, R. (1999) Development 126, 20332044.
(5) Brückner, K. et al. (1997) Science 275, 1640-1643.
(6) Palmer, A. et al. (2002) Mol. Cell 9, 725-737.
Specificity/Sensitivity: EphA2 (D4A2) XP® Rabbit mAb
recognizes endogenous levels of total EphA2 protein.
Source/Purification: Monoclonal antibody is produced
by immunizing animals with a human EphA2 recombinant
protein fragment.
**Anti-rabbit secondary antibodies must be used to
detect this antibody.
m
CO
9
B1
C6
kDa
SN
Background: The Eph receptors are the largest known family of receptor tyrosine kinases (RTKs). They can be divided
into two groups based on sequence similarity and on their
preference for a subset of ligands: EphA receptors bind to a
glycosylphosphatidylinositol-anchored ephrin A ligand, and
EphB receptors bind to ephrin B proteins that have a transmembrane and cytoplasmic domain (1,2). Eph receptors and
ligands may be involved in many diseases including cancer
(3). Both ephrin A and ephrin B ligands have dual functions.
As RTK ligands, the ephrins stimulate the kinase activity
of the Eph receptors and activate signaling pathways in
receptor-expressing cells. The ephrin extracellular domain
is sufficient for this function as long as it is clustered
(4). The second function of ephrins has been described
as “reverse signaling,” whereby the cytoplasmic domain
becomes tyrosine phosphorylated, allowing interactions
with other proteins that may activate signaling pathways in
the ligand-expressing cells (5). Various stimuli can induce
tyrosine phosphorylation of ephrin B, including binding to
EphB receptors, activation of Src kinase and stimulation by
PDGF and FGF (6). Tyrosines 324/327 have been identified
as major phosphorylation sites of ephrin B1 in vivo (7).
(7) Kalo, M.S. et al. (2001) J. Biol. Chem. 276, 38940-38948.
(8) Guo, H. et al. (2006) Cancer Res 66, 7050-7058.
SK-MEL-28
IMPORTANT: For western blots, incubate membrane
with diluted antibody in 5% w/v BSA, 1X TBS, 0.1%
Tween® 20 at 4°C with gentle shaking, overnight.
Confocal immunofluorescent analysis of A549 cells (upper)
and SK-MEL-28 cells (lower) using EphA2 (D4A2) XP® Rabbit
mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent
DNA dye).
Tween® is a registered trademark of ICI Americas, Inc.
DRAQ5® is a registered trademark of Biostatus Limited.
Applications Key:
W—Western
Species Cross-Reactivity Key:
IP—Immunoprecipitation
H—human
M—mouse
Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans
IHC—Immunohistochemistry
R—rat
Hr—horse
Hm—hamster
ChIP—Chromatin Immunoprecipitation
Mk—monkey
Mi—mink
All—all species expected
C—chicken
IF—Immunofluorescence
F—Flow cytometry
Dm—D. melanogaster X—Xenopus
Z—zebrafish
E-P—ELISA-Peptide
B—bovine
Species enclosed in parentheses are predicted to react based on 100% homology.
Immunohistochemical analysis of paraffin-embedded SNB19 (left) and SKMEL28 (right) cell pellets using EphA2 (D4A2) XP® Rabbit
mAb.
Immunohistochemical analysis of paraffin-embedded ovarian
carcinoma using EphA2 (D4A2) XP® Rabbit mAb.
© 2014 Cell Signaling Technology, Inc.
Immunohistochemical analysis of paraffin-embedded human
non-small cell lung carcinoma using EphA2 (D4A2) XP® Rabbit
mAb.
Orders n 877-616-CELL (2355)
[email protected]
Support n 877-678-TECH (8324)
[email protected]
Web n www.cellsignal.com