Revision 6

#7034

Store at +4C

PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H

UniProt ID:
#P07949

Entrez-Gene Id:
#5979

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
Ret Rabbit Monoclonal Antibody Coated Microwells6250196 tests+4C
Phospho Tyrosine Mouse Detection Monoclonal Antibody129821 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

The PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ret protein. A Ret rabbit mAb has been coated on the microwells. After incubation with cell lysates, Ret protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse mAb is added to detect captured tyrosine-phosphorylated Ret protein. 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 the absorbance for this developed color is proportional to the quantity of Ret protein phosphorylated on tyrosine residues.

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

Specificity/Sensitivity

PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit #7034 detects endogenous levels of tyrosine-phosphorylated Ret 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

The Ret proto-oncogene (c-Ret) is a receptor tyrosine kinase that functions as a multicomponent receptor complex in conjunction with other membrane-bound, ligand-binding GDNF family receptors (1). Ligands that bind the Ret receptor include the glial cell line-derived neurotrophic factor (GDNF) and its congeners neurturin, persephin, and artemin (2-4). Research studies have shown that alterations in the corresponding RET gene are associated with diseases including papillary thyroid carcinoma, multiple endocrine neoplasia (type 2A and 2B), familial medullary thyroid carcinoma, and a congenital developmental disorder known as Hirschsprung’s disease (1,3). The Tyr905 residue located in the Ret kinase domain plays a crucial role in Ret catalytic and biological activity. Substitution of Phe for Tyr at position 905 dramatically inhibits Ret autophosphorylation activity (5).

Trademarks and Patents

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

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

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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

使用に関する制限

使用に関する制限

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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ロゴ
Figure 1: Constitutive phosphorylation of Ret in TT cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit #7034 (upper, right). In contrast, a low level of phospho-Ret protein is detected in TT cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Ret protein from both nonphospho or phospho lysates are detected by sandwich ELISA (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-Ret (Tyr905) Rabbit Antibody #3221 (right) or a total Ret (C31B4) Rabbit mAb #3223 (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.
ELISA and/or ELISA-like Assays Image 1: PathScan<sup>®</sup> Phospho-Ret (panTyr) Sandwich ELISA Kit
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. TT cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate).
ELISA and/or ELISA-like Assays Image 2: PathScan<sup>®</sup> Phospho-Ret (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.