Axl (C89E7) Rabbit mAb - Cell Signaling Technology · PDF file#8661 Store at -20°C Axl...
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#8661 Store at -20°C Axl (C89E7) Rabbit mAb W, IP, IHC-P, IF-IC, F Endogenous H, Mk 138 kDa Rabbit IgG** Background: Axl, Sky, and Mer are three members of a receptor tyrosine kinase (RTK) family that share a conserved intracellular tyrosine kinase domain and an extracellular domain similar to those seen in cell adhesion molecules. These RTKs bind the vitamin K-dependent protein growth- arrest-specific 6 (Gas6), which is structurally related to the protein S anticoagulation factor (1). Upon binding to its receptor, Gas6 activates phosphatidylinositol 3-kinase (PI3K) and its downstream targets Akt and S6K, as well as NF-κB (2,3). A large body of evidence supports a role for Gas6/Axl signaling in cell growth and survival in normal and cancer cells (4). Specificity/Sensitivity: Axl (C89E7) Rabbit mAb detects endogenous levels of total Axl protein. Source/Purification: Monoclonal antibody is produced by immunizing animals with a recombinant human fragment of the Axl protein. 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. *Species cross-reactivity is determined by western blot. **Anti-rabbit secondary antibodies must be used to detect this antibody. Recommended Antibody Dilutions: Western blotting 1:1000 Immunoprecipitation 1:50 Immunohistochemistry (Paraffin) 1:600† Unmasking buffer: Citrate 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 Flow Cytometry 1:100 For product specific protocols please see the web page for this product at www.cellsignal.com. Please visit www.cellsignal.com for a complete listing of recommended companion products. Background References: (1) Crosier, K.E. and Crosier, P.S. (1997) Pathology 29, 131-135. (2) Demarchi, F. et al. (2001) J. Biol. Chem. 276, 31738-31744. (3) Lee, W. P. et al. (2002) Oncogene 21, 329-336. (4) Bellosta, P. et al. (1997) Oncogene 15, 2387-2397. Orders n 877-616-CELL (2355) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com Applications Species Cross-Reactivity* Molecular Wt. Isotype Entrez-Gene ID #558 UniProt ID #P30530 Species Cross-Reactivity Key: H—human M—mouse R—rat Hm—hamster Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebrafish B—bovine Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans Hr—horse All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology. Applications Key: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide IMPORTANT: For western blots, incubate mem- brane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween ® 20 at 4°C with gentle shaking, overnight. ® 2014 Cell Signaling Technology, Inc. SignalStain ® and Cell Signaling Technology ® are trademarks of Cell Signaling Technology, Inc. Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Confocal immunofluorescent analysis of DU 145 (upper) and HCC827 (lower) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5 ® #4084 (fluorescent DNA dye). DRAQ5 ® is a registered trademark of Biostatus Limited. U.S. Patent No. 5,675,063 DU 145 HCC827 Events Axl Flow cytometric analysis of Jurkat cells (blue) and DU145 cells (green) using Axl (C89E7) Rabbit mAb. kDa Axl β-Actin H1255 HeLa H2347 DU 145 HCC827 Jurkat COS-7 200 140 100 80 60 50 40 30 60 50 40 Tween ® is a registered trademark of ICI Americas, Inc. For Research Use Only. Not For Use In Diagnostic Procedures. rev. 03/20/17
Transcript of Axl (C89E7) Rabbit mAb - Cell Signaling Technology · PDF file#8661 Store at -20°C Axl...
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Axl (C89E7) Rabbit mAb
W, IP, IHC-P, IF-IC, FEndogenous
H, Mk 138 kDa Rabbit IgG**
Background: Axl, Sky, and Mer are three members of a receptor tyrosine kinase (RTK) family that share a conserved intracellular tyrosine kinase domain and an extracellular domain similar to those seen in cell adhesion molecules. These RTKs bind the vitamin K-dependent protein growth-arrest-specific 6 (Gas6), which is structurally related to the protein S anticoagulation factor (1). Upon binding to its receptor, Gas6 activates phosphatidylinositol 3-kinase (PI3K) and its downstream targets Akt and S6K, as well as NF-κB (2,3). A large body of evidence supports a role for Gas6/Axl signaling in cell growth and survival in normal and cancer cells (4).
Specificity/Sensitivity: Axl (C89E7) Rabbit mAb detects endogenous levels of total Axl protein.
Source/Purification: Monoclonal antibody is produced by immunizing animals with a recombinant human fragment of the Axl protein.
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.*Species cross-reactivity is determined by western blot.
** Anti-rabbit secondary antibodies must be used to detect this antibody.
Recommended Antibody Dilutions: Western blotting 1:1000 Immunoprecipitation 1:50 Immunohistochemistry (Paraffin) 1:600† Unmasking buffer: Citrate 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 Flow Cytometry 1:100
For product specific protocols please see the web page for this product at www.cellsignal.com.Please visit www.cellsignal.com for a complete listing of recommended companion products.
Background References: (1) Crosier, K.E. and Crosier, P.S. (1997) Pathology 29,
131-135.
(2) Demarchi, F. et al. (2001) J. Biol. Chem. 276, 31738-31744.
(3) Lee, W. P. et al. (2002) Oncogene 21, 329-336.
(4) Bellosta, P. et al. (1997) Oncogene 15, 2387-2397.
Orders n 877-616-CELL (2355)[email protected]
Support n 877-678-TECH (8324)[email protected]
Web n www.cellsignal.com
Applications Species Cross-Reactivity* Molecular Wt. Isotype
Entrez-Gene ID #558 UniProt ID #P30530
Species Cross-Reactivity Key: H—human M—mouse R—rat Hm—hamster Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebrafish B—bovine
Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans Hr—horse All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology.
Applications Key: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide
IMPORTANT: For western blots, incubate mem-brane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
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Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of DU 145 (upper) and HCC827 (lower) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
DRAQ5® is a registered trademark of Biostatus Limited.
U.S. Patent No. 5,675,063
DU 145
HCC827
Even
ts
AxlFlow cytometric analysis of Jurkat cells (blue) and DU145 cells (green) using Axl (C89E7) Rabbit mAb.
kDa
Axl
β-Actin
H1255
HeLa
H2347
DU 145
HCC827
Jurka
tCOS-7
20014010080
60
50
40
30
60
5040
Tween® is a registered trademark of ICI Americas, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.
rev. 03/20/17
Orders n 877-616-CELL (2355) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com® 2
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, Inc
.
Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
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