Revision 5
#7091
Store at +4C
PathScan® Total ROS1 Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H
UniProt ID:
#P08922
Entrez-Gene Id:
#6098
877-616-CELL (2355)
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 |
|---|---|---|---|---|
| ROS1 Mouse mAb Coated Microwells | 30828 | 96 tests | +4C | |
| ROS1 Rabbit Detection mAb | 13746 | 1 ea | Green (Lyophilized) | +4C |
| Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) | 13272 | 1 ea | Red (Lyophilized) | +4C |
| Detection Antibody Diluent | 13339 | 11 ml | Green | +4C |
| HRP Diluent | 13515 | 11 ml | Red | +4C |
| TMB Substrate | 7004 | 11 ml | +4C | |
| STOP Solution | 7002 | 11 ml | +4C | |
| Sealing Tape | 54503 | 2 ea | +4C | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | +4C | |
| ELISA Sample Diluent | 11083 | 25 ml | Blue | +4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml | -20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Total ROS1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of ROS1 protein. A ROS1 mouse antibody has been coated onto the microwells. After incubation with cell lysates, ROS1 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a ROS1 rabbit detection antibody is added to detect captured ROS1 protein. Anti-rabbit 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 ROS1 protein.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
PathScan® Total ROS1 Sandwich ELISA Kit #7091 detects endogenous levels of ROS1 protein in human cells, as shown in Figure 1. The 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
ROS1, an orphan receptor tyrosine kinase of the insulin receptor family, was initially identified as a homolog of v-ros from the UR2 sarcoma virus (1). ROS1 consists of a large extracellular domain that is composed of six fibronectin repeats, a transmembrane domain, and a C-terminal kinase domain. Being an orphan receptor, the functions of ROS1 are not well known, though it has been shown to play an important role in differentiation of epididymal epithelium (2). The first oncogenic fusion of ROS1, FIG-ROS1, was initially identified by research studies in glioblastoma (3), and subsequent studies have found this fusion in cholangiocarcinoma (4), ovarian cancer (5), and non-small cell lung cancer (NSCLC) (6). Investigators have found additional oncogenic ROS1 fusion proteins in NSCLC (at a frequency of ~1.6%), where the ROS1 kinase domain is fused to the amino-terminal region of several different proteins, including CD74 and SLC34A2 (6-8). ROS1 fusion proteins activate the SHP-2 phosphatase, PI3K/Akt/mTOR, Erk, and Stat3 pathways (3,4,9). There are two autophosphorylation sites (Tyr2274, Tyr2334) downstream of the kinase domain of ROS1, either of which may serve as biomarkers of ROS1 kinase activity, including that of ROS1 fusion proteins (10).
Background References
- Matsushime, H. et al. (1986) Mol Cell Biol 6, 3000-4.
- Yeung, C.H. et al. (1999) Biol Reprod 61, 1062-9.
- Charest, A. et al. (2003) Genes Chromosomes Cancer 37, 58-71.
- Gu, T.L. et al. (2011) PLoS One 6, e15640.
- Birch, A.H. et al. (2011) PLoS One 6, e28250.
- Rimkunas, V.M. et al. (2012) Clin Cancer Res 18, 4449-57.
- Rikova, K. et al. (2007) Cell 131, 1190-203.
- Stumpfova, M. and Jänne, P.A. (2012) Clin Cancer Res 18, 4222-4.
- Jun, H.J. et al. (2012) Cancer Res 72, 3764-74.
- Zou, H.Y. et al. (2015) Proc Natl Acad Sci U S A 112, 3493-8.
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus 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 GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
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.
Revision 5
Figure 1: Constitutive phosphorylation of ROS1 in HCC78 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-ROS1 (panTyr) Sandwich ELISA Kit #7093 (upper, right). In contrast, a low level of phospho-ROS1 protein is detected in HCC78 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total HCC78 protein from both nonphospho and phospho lysates are detected by PathScan® Total ROS1 Sandwich ELISA Kit #7091 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-ROS1 (Tyr2274) Antibody #3078 (right) or total ROS1 (69D6) Mouse mAb #3266 (left) are shown in the bottom figure.
*Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and Na3VO4.
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. HCC78 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate, respectively).
