{"product_id":"revvity-64akspet","title":"Revvity, 64AKSPET, HTRF Human Phospho-AKT (Ser473) Detection Kit, 96 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThe Phospho-AKT (Ser473) kit is designed to robustly and efficiently monitor AKT phosphorylation on Ser473 across all isoforms, as a readout of Pi3K pathway activation. Also known as Protein Kinase B (PKB), AKT is an oncogene that controls apoptosis, cell proliferation, transcription, cell migration, and glucose metabolism.\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-AKT (Ser473) assay principle\u003cbr\u003e\nThe Phospho-AKT (Ser473) assay measures AKT when phosphorylated at Ser473. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-AKT (Ser473) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one 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 independent 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 proteins 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-AKT (Ser473) 2-plate assay protocol\u003cbr\u003e\nThe 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-AKT (Ser473) assay 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-AKT (Ser473) assay 1-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated AKT (Ser473)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\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\nUnit Size-96 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775236231337,"sku":"64AKSPET","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64akspet","provider":"Iright","version":"1.0","type":"link"}