Revision 6

#18350

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit Monoclonal Antibody4446320 µl110-130, 125-140, 180-200 kDaRabbit IgG
Phospho-Axl (Tyr702) (D12B2) Rabbit Monoclonal Antibody572420 µl138 kDaRabbit IgG
Axl (C89E7) Rabbit Monoclonal Antibody866120 µl138 kDaRabbit IgG
Mer (D21F11) Rabbit Monoclonal Antibody431920 µl210 kDaRabbit IgG
Tyro3 (D38C6) Rabbit Monoclonal Antibody558520 µl110, 130 kDaRabbit IgG
GAS6 (D3A3G) Rabbit Monoclonal Antibody6720220 µl78 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Tyro/Axl/Mer Activation Antibody Sampler Kit provides an economical means of detecting the activation of TAM family members using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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.

Background

Axl, Mer and Tyro3 are three members of the TAM family receptor tyrosine kinase that share a common NCAM (neural adhesion molecule)-related extracellular domain and a conserved intracellular tyrosine kinase domain. These receptors bind common homologous vitamin K dependent protein GAS6 and protein S to activate downstream signaling pathways (1). TAM family receptors are involved in the development of immune, nervous, vascular and reproductive systems, autoimmune disease, cancer drug resistance and tumor immunity response (2-5). Axl (Tyr698), Axl (Tyr702), Mer Tyr(749) and Tyro3 (Tyr681) are conserved autophosphorylation sites located in the activation loop of the respective tyrosine kinase domains. Phosphorylation at these sites is required for full kinase activation of each of the corresponding receptors (6,7).

Trademarks and Patents

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

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Western blot analysis of extracts from SK-MEL-5, Hep G2 and Jurkat cells using Mer (D21F11) XP® Rabbit mAb.
undefined Image 1: Tyro/Axl/Mer Activation Antibody Sampler Kit
Western blot analysis of extracts from NCI-H1299 and SK-MEL-5 cells, untreated (-) or treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min, using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) (D6M4W) Rabbit mAb (upper) and Axl (C89E7) Rabbit mAb #8661 (lower 1), Mer (D21F11) XP® Rabbit mAb #4319 (lower 2), and Tyro3 (D38C6) Rabbit mAb #5585 (lower 3).
undefined Image 2: Tyro/Axl/Mer Activation Antibody Sampler Kit
Western blot analysis of extracts from COS-7 cells, and tissues from human and mouse brain using Tyro3 (D38C6) Rabbit mAb.
undefined Image 3: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Western blot analysis of extracts from NCI-H1299 cells, untreated or Gas6-treated (100 ng/ml for 10 minutes), using Phospho-Axl (Tyr702) (D12B2) Rabbit mAb (upper) or Axl (C44G1) Rabbit mAb #4566 (lower).
undefined Image 4: Tyro/Axl/Mer Activation Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using GAS6 (D3A3G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 5: Tyro/Axl/Mer Activation Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 6: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Western blot analysis of HeLa Cell Extracts, untreated (-) or Axl knock-out (+) using Axl (C89E7) Rabbit mAb, #8661 (upper) or GAPDH (D16H11) XP® Rabbit mAb, #5174 (lower).
undefined Image 7: Tyro/Axl/Mer Activation Antibody Sampler Kit
Confocal immunofluorescent analysis of SK-MEL-5 cells (left), Hep G2 cells (center) and MCF7 cells (right) using Mer (D21F11) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ® #4084 (fluorescent DNA dye).
undefined Image 8: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunoprecipitation of Phospho-Axl (Tyr698) protein from NCI-H1299 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Axl (C89E7) Rabbit mAb #8661. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Axl (C89E7) Rabbit mAb #8661 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
undefined Image 9: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GAS6 (D3A3G) Rabbit mAb.
undefined Image 10: Tyro/Axl/Mer Activation Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Axl (C89E7) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 11: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunoprecipitation of Phospho-Mer (Tyr749) protein from SK-MEL-5 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Mer(D21F11) XP® Rabbit mAb #4319. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Mer (D21F11) XP® Rabbit mAb #4319 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
undefined Image 12: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded NCI-H226 cell pellet (left, positive) or HT29 cell pellet (right, negative) using GAS6 (D3A3G) Rabbit mAb.
undefined Image 13: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded breast carcinoma using Axl (C89E7) Rabbit mAb. Note staining of infiltrating cells.
undefined Image 14: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunoprecipitation of Phospho-Tyro3 (Tyr681) protein from SK-MEL-5 cells treated with GAS6 (400 ng/ml) and H2O2 (4 mM) for 10 min. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lane 3 is Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb, and lane 4 is Tyro 3 (D38C6) Rabbit mAb #5585. Western blot analysis was performed using Phospho-Axl (Tyr698)/Mer (Tyr749)/Tyro3 (Tyr681) Rabbit mAb (upper) and Tyro3 (D38C6) Rabbit mAb #5585 (lower). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
undefined Image 15: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using GAS6 (D3A3G) Rabbit mAb.
undefined Image 16: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using Axl (C89E7) Rabbit mAb.
undefined Image 17: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human serous carcinoma of the ovary using GAS6 (D3A3G) Rabbit mAb.
undefined Image 18: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using Axl (C89E7) Rabbit mAb.
undefined Image 19: Tyro/Axl/Mer Activation Antibody Sampler Kit
Confocal immunofluorescent analysis of NCI-H226 (left, positive) and HT-29 (right, negative) using GAS6 (D3A3G) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 20: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using Axl (C89E7) Rabbit mAb.
undefined Image 21: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded metastatic lung carcinoma using Axl (C89E7) Rabbit mAb.
undefined Image 22: Tyro/Axl/Mer Activation Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded cell pellets, NCI-H1299 (left) or Jurkat (right), using Axl (C89E7) Rabbit mAb.
undefined Image 23: Tyro/Axl/Mer Activation Antibody Sampler Kit
Confocal immunofluorescent analysis of DU 145 (left) and HCC827 (right) cells using Axl (C89E7) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 24: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Flow cytometric analysis of fixed and permeabilized Jurkat cells (blue, negative) and DU145 cells (green, positive) using Axl (C89E7H4) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 25: Tyro/Axl/Mer Activation Antibody Sampler Kit
Flow cytometric analysis of MCF7 cells (blue) and HepG2 cells (green) using Mer (D21F11) XP® Rabbit mAb (solid lines) or a concetration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 26: Tyro/Axl/Mer Activation Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using Axl (C89E7) Rabbit mAb #8661. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 27: Tyro/Axl/Mer Activation Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.