Revision 7

#53402

Store at +4C

PathScan® Phospho-Tyro3 (panTyr) Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H

UniProt ID:
#Q06418

Entrez-Gene Id:
#7301

CST Logo
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 Color Storage Temp
Tyro3 Rabbit mAb Coated Microwells6956896 tests+4C
Phospho Tyrosine Mouse Detection mAb129821 eaGreen (Lyophilized)+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)133041 eaRed (Lyophilized)+4C
Detection Antibody Diluent1333911 mlGreen+4C
HRP Diluent1351511 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
ELISA Sample Diluent1108325 mlBlue+4C
Cell Lysis Buffer (10X)980315 ml-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

PathScan® Phospho-Tyro3 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Mer protein. A Tyro3 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Tyro3 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Tyrosine Mouse Detection mAb is added to detect the captured phospho-Tyro3 proteins. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of Tyro3 protein phosphorylated on tyrosine residues.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Phospho-Tyro3 (panTyr) Sandwich ELISA Kit recognizes endogenous levels of tyrosine-phosphorylated Tyro3 protein in human cells, as shown in Figure 1. This kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Tyro3 is a receptor tyrosine kinase belonging to the TAM subfamily (Tyro3, Axl, and Mer). All three members have similar domain structures composed of an extracellular region with 2 Ig-like domains, followed by 2 FNII-like domains, a single transmembrane region, and a cytoplasmic tyrosine kinase domain (1). The natural ligand for Tyro3, as well as Axl and Mer, is GAS6 (growth arrest specific gene 6) (1,2). Expression pattern and target knockout data indicate an important role of Tyro3 in apoptotic cell phagocytosis of dendritic cells and macrophages (3), NK cell differentiation (4), reproductive neuron survival and migration (5), osteoclast stimulation (6,7), as well as cortical and hippocampal neuron function (8). Both MAPK and PI3K pathways have been suggested as downstream targets of Tyro3 activation (7,8). Tyro3 has also been shown to be correlated to melanoma tumorigenesis, likely through its regulatory role in the expression of oncogenic microphthalmia-associated transcription factor (MITF) (9).

Trademarks and Patents

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

PathScan is a registered 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 7

Cell Signaling Technology Logo
Figure 1. Constitutive phosphorylation of Tyro3 in CHO/Tyro3 cells lysed in the presence of phosphatase inhibitors (phospho lysate) is detected by PathScan® Phospho-Tyro3 (panTyr) Sandwich ELISA Kit. In contrast, a low level of phospho-Tyro3 protein is detected in CHO/Tyro3 cells lysed in the absence of phosphatase inhibitors (nonphospho lysate). Similar levels of total Tyro3 protein from both nonphospho and phospho lysates are detected by PathScan® Total Tyro3 Sandwich ELISA Kit #39412 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Tyro3(D38C6) Rabbit mAb #5585 (left) and a Phospho-Tyrosine Mouse mAb #9411 (right) are shown in the bottom figure. Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and sodium orthovanadate.

CHO/Tyro3 cells are derived from CHO cells tranfected with human Tyro3.

ELISA Image 1: PathScan<sup>®</sup> Phospho-Tyro3 (panTyr) Sandwich ELISA Kit
Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm as detected by the PathScan® Phospho-Tyro3 (panTyr) Sandwich ELISA Kit is shown. Unstarved CHO cells were cultured (85% confluence) and transfected with human Tyro3 for 6 hrs, then lysed with or without addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).
ELISA Image 2: PathScan<sup>®</sup> Phospho-Tyro3 (panTyr) Sandwich ELISA 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.