{"product_id":"revvity-64atg14s9peg","title":"Revvity, 64ATG14S9PEG, HTRF Human and Mouse Phospho-ATG14 (Ser29) Detection Kit, 500 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nATG14, Autophagy related protein 14, or BAKOR for Beclin 1-associated autophagy-related key regulator, is a key player in the autophagosome nucleation step in macroautophagy. Upon cellular stress, the nutrient\/energy-sensitive sensors mTOR and AMPK lead to the activation of the ULK1 complex, which allows phosphorylation of ATG14 on Serine 29, in turn enabling the downstream activation of the PIK3C3 autophagosome nucleation complex.\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\nHTRF Phospho ATG14 assay principle\u003cbr\u003e\nThe HTRF Phospho-ATG14 (Ser29) assay measures ATG14 when phosphorylated at Ser 29. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Phospho-ATG14 (Ser29) assay uses 2 labeled antibodies: one with a donor fluorophore, the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving the two labeled antibodies 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\u003eHTRF Phospho-ATG14 Ser29 2-plate assay protocol\u003cbr\u003e\nThe 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of the Phospho-ATG14 Ser 29 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\u003eHTRF Phospho-ATG14 Ser29 one-plate assay protocol\u003cbr\u003e\nDetection of phospho ATG14 Ser 29 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\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 Mouse\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nUnit Size-500 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775262445737,"sku":"64ATG14S9PEG","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64atg14s9peg","provider":"Iright","version":"1.0","type":"link"}