Revision 5

#7980Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

B

UniProt ID:

#P29474

Entrez-Gene Id:

#4846

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Phospho-eNOS Rabbit mAb Coated Microwells 55732 96 tests +4C
eNOS Mouse Detection mAb 49999 1 ea Green (Lyophilized) +4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 13304 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® Phospho-eNOS (Ser1177) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of eNOS when phosphorylated at Ser1177. A phospho-eNOS (Ser1177) rabbit monoclonal antibody has been coated onto the microwells. After incubation with cell lysates, phospho-eNOS protein is captured by the coated antibody. Following extensive washing, an eNOS mouse monoclonal detection antibody is added to detect captured eNOS protein phosphorylated at Ser1177. HRP-linked streptavidin 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 eNOS phosphorylated at Ser1177.

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

Specificity/Sensitivity

PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit #7980 detects endogenous levels of eNOS protein only when phosphorylated at Ser1177 (see 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

Endothelial nitric-oxide synthase (eNOS) is an important enzyme in the cardiovascular system. It catalyzes the production of nitric oxide (NO), a key regulator of blood pressure, vascular remodeling, and angiogenesis (1,2). The activity of eNOS is regulated by phosphorylation at multiple sites. The two most thoroughly studied sites are the activation site Ser1177 and the inhibitory site Thr495 (3). Several protein kinases including Akt/PKB, PKA, and AMPK activate eNOS by phosphorylating Ser1177 in response to various stimuli (4,5). In contrast, bradykinin and H2O2 activate eNOS activity by promoting both Ser1177 phosphorylation and Thr495 dephosphorylation (6,7).

  1. Fulton, D. et al. (2001) J Pharmacol Exp Ther 299, 818-24.
  2. Shaul, P.W. (2002) Annu Rev Physiol 64, 749-74.
  3. Chen, Z.P. et al. (1999) FEBS Lett 443, 285-9.
  4. Dimmeler, S. et al. (1999) Nature 399, 601-5.
  5. Fulton, D. et al. (1999) Nature 399, 597-601.
  6. Harris, M.B. et al. (2001) J Biol Chem 276, 16587-91.
  7. Thomas, S.R. et al. (2002) J Biol Chem 277, 6017-24.

Background References

    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 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.
    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.

    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
    #7980

    PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit

    PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. Treatment of BAEC cells with 500 µM of H2O2 increases phosphorylation of eNOS at Ser1177, detected by the PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit #7980. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using eNOS Antibody #9572 (left panel) or Phospho-eNOS (Ser1177) (C9C3) Rabbit mAb #9570 (right panel) are shown in the bottom figure.
    PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit: Image 2 Expand Image
    Figure 2. The relationship between the protein concentration of lysates from BAEC cells treated with H2O2 or λ-phosphatase-treated lysate and the absorbance at 450 nm using PathScan® Phospho-eNOS (Ser1177) Sandwich ELISA Kit #7980 is shown.