Product Pathways - PathScan ELISA
PathScan® Total iNOS Sandwich ELISA Kit #7097
| Kit Includes | Volume | Solution Color |
|---|---|---|
| iNOS Ab Coated Microwells | 96 tests | |
| iNOS Detection Ab | 11 milliliters | Green |
| Anti-Mouse IgG HRP-linked Antibody | 11 milliliters | Red |
| TMB Substrate | 11 milliliters | Colorless |
| STOP Solution | 11 milliliters | Colorless |
| Sealing Tape | 2 sheets | |
| Wash Buffer (20X) | 25 milliliters | Colorless |
| Sample Diluent | 25 milliliters | Blue |
| Cell Lysis Buffer (10X) # 9803 | 15 milliliters | Yellowish |
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer (10X), which is stored at –20°C (packaged separately).
Species Cross-Reactivity
M
Reactivity Key: M=Mouse
Description
The PathScan® Total iNOS Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of iNOS. An iNOS rabbit antibody has been coated onto the microwells. After incubation with cell lysates, iNOS protein is captured by the coated antibody. Following extensive washing, an iNOS mouse detection antibody is added to detect the captured iNOS protein. Anti-mouse IgG, HRP-linked Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of iNOS.
Specificity / Sensitivity
Cell Signaling Technology's PathScan® Total iNOS Sandwich ELISA Kit #7097 detects endogenous levels of total iNOS protein.
ELISA
Figure 2. The relationship between the protein concentration of lysates from untreated and LPS-treated Raw264.7 cells and the absorbance at 450 nm is shown. Raw264.7 cells (80-90% confluent) were treated with 1 μg/ml LPS for 6 hours at 37ºC and then lysed.
ELISA - Western correlation
Figure 1. Treatment of Raw264.7 cells with LPS stimulates expression of iNOS, detected by the PathScan® Total iNOS Sandwich ELISA Kit. Raw264.7 cells (80-90% confluent) were treated with 1 ng/ml LPS for 6 hours at 37º C. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using iNOS Antibody (Mouse Specific) #2982 is shown in the bottom figure.
Background
Nitric Oxide Synthase (NOS) catalyses the formation of nitric oxide (NO) and citruline from L-arginine, oxygen and cofactors. Three family members have been characterized: neuronal NOS (nNOS), which is found primarily in neuronal tissue; inducible NOS (iNOS), which is induced by interferon gamma and lipopolysaccharides in the kidney and cardiovascular system; and endothelial NOS (eNOS), which is expressed in blood vessels (1). NO is a messenger molecule with diverse functions throughout the body including the maintenance of vascular integrity, homeostasis, synaptic plasticity, long-term potentiation, learning, and memory (2,3).
- Tsutsui, M. (2004) J Atheroscler Thromb 11, 41-48.
- Son, H. et al. (1996) Cell 87, 1015-10123.
- Hawkins, R.D. (1996) Neuron 16, 465-467.
Application References
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Companion Products
- 2982 iNOS Antibody (Mouse Specific)
- 2977 iNOS Antibody
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
