{"product_id":"revvity-64enoss7pey","title":"Revvity, 64ENOSS7PEY, HTRF Human Phospho-eNOS (Ser1177) Detection Kit, 50,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis HTRF cell-based assay conveniently and accurately detects phosphorylated eNOS at Serine 1177.\u003cbr\u003e\neNOS is expressed primarily in endothelial cells, where its function is to control blood vessel dilation, blood pressure, and numerous other anti-atherosclerotic and vasoprotective effects.\u003cbr\u003e\nSerine 1177 is the most important phosphorylation site of eNOS, positively modulating its activity. Phosphorylation at serine 1177 is under the control of several kinases, including AKT, PKA, AMPK, CaMKK2b, and CHKI\u003cbr\u003e\nThis HTRF cell-based assay conveniently and accurately detects phosphorylated eNOS at Serine 1177.\u003cbr\u003e\n​​​​​​​​​​​​​​\u003cbr\u003e\nHTRF assays offer many advantages over other technologies:\u003c\/p\u003e\n\n\u003cp\u003eHomogeneous add-and-read format\u003cbr\u003e\nNo wash steps\u003cbr\u003e\nLow background\u003cbr\u003e\nStraightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates\u003cbr\u003e\nStable signal, providing flexibility in time of readout or size of assays\u003cbr\u003e\nHow it works\u003cbr\u003e\nPhospho-eNOS (Ser1177) assay principle\u003cbr\u003e\nThe Phospho-eNOS (S1177) assay measures eNOS when phosphorylated at serine 1177. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses 2 antibodies, one labeled with a donor fluorophore and the other with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies, and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein's phosphorylation state under a no-wash assay format.\u003c\/p\u003e\n\n\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-eNOS (Ser1177) two-plate assay protocol\u003cbr\u003e\nThe two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of Phospho-eNOS (Ser1177) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.\u003c\/p\u003e\n\n\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-eNOS (Ser1177) one-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated eNOS (Ser1177) with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nLysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4 Lysis Buffer 5\u003cbr\u003e\nMolecular Modification-Phosphorylation\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nSample Volume-16 µL\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Cardiovascular Metabolism\/Diabetes\u003cbr\u003e\nUnit Size-50,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775246749865,"sku":"64ENOSS7PEY","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/ar\/products\/revvity-64enoss7pey","provider":"Iright","version":"1.0","type":"link"}