The Sphingosine 1-Phosphate ELISA kit (S1P ELISA) is a sensitive and robust method for the quantification of S1P in biological samples (96-well format).
Assay Range: 0.0625 µM – 2 µM
Sample Volume: For duplicate data points 25 µL of each sample is required.
Sample Type: This kit detects S1P from human or animal in serum, plasma, tissue homogenate, and cell lysate, This assay is not species specific and has been tested with S1P from human, bovine, equine, caprine, and mouse.
The Sphingosine 1-Phosphate ELISA kit (S1P ELISA) measures S1P in biological samples.
To run the S1P ELISA, the S1P coated plate is first blocked to reduce non-specific binding. Then the S1P standards and samples are combined with an anti-S1P antibody before it is transferred to the blocked S1P coated plate. The S1P coated plate and the S1P in the sample compete for binding of the anti-S1P antibody during this step. Following an incubation and plate wash, streptavidin-HRP is added to the plate for detection of any bound anti-S1P antibodies (labeled with biotin). After an additional incubation and plate wash, TMB substrate is added to the plate. The colorometric TMB reaction is stopped with the addition of 1N sulfuric acid and the plate is measured at 450 nm absorbance. The concentration of S1P, in the samples, is determined when compared to the S1P standard curve.
Sphingosine 1-Phosphate (S1P) is a key component of the sphingolipid signaling cascade. S1P initiates a proliferative, pro-angiogenic and anti-apoptotic sequence of events contributing to cancer progression. Recently, scientific literature has suggested that S1P is a potent tumorigenic growth factor that is likely released from tumor cells and that S1P may be a novel biomarker for early stage cancer detection. Sphingosine kinase has also been shown to be up-regulated in a variety of cancer types (S1P is produced via the activity of sphingosine kinase phosphorylating sphingosine).
Detection of S1P Lyase activity is also possible with this assay using a modified protocol. See below for the protocol TDS.
1. Lai, W. Q., A. W. Irwan, et al. (2008). “Anti-inflammatory effects of sphingosine kinase modulation in inflammatory arthritis.” J Immunol 181(11): 8010-7.
2. Kirby, R. J., Y. Jin, et al. (2009). “Dynamic regulation of sphingosine-1-phosphate homeostasis during development of mouse metanephric kidney.” Am J Physiol Renal Physiol 296(3): F634-41.
3. Nincheri, P., C. Bernacchioni, et al. (2010). “Sphingosine kinase-1/S1P1 signalling axis negatively regulates mitogenic response elicited by PDGF in mouse myoblasts.” Cell Signal 22(11): 1688-99.
4. Roviezzo, F., V. Brancaleone, et al. (2011). “Sphingosine-1-Phosphate Modulates Vascular Permeability and Cell Recruitment in Acute Inflammation In Vivo.” Journal of Pharmacology and Experimental Therapeutics 337(3): 830-837.
Note to purchaser: The S1P Assay was co-developed using a proprietary monoclonal antibody from Lpath Inc. (San Diego, CA), now Apollo Endosurgery (Austin, TX). Lpath, Inc. has a number of patent applications pending or issued covering material(s) incorporated into this assay. The assay`s intended use is for the measurement of S1P in biological samples. Any other use, including drug discovery, requires a license from Lpath (www.lpath.com).