{"product_id":"revvity-64atg14tpeh","title":"Revvity, 64ATG14TPEH, HTRF Human Total ATG14 Detection Kit, 10,000 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\nTotal ATG14 assay principle\u003cbr\u003e\nThe Total-ATG14 assay quantifies the expression level of ATG14 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total-ATG14 assay uses two labeled antibodies, one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of ATG14 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression 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\n\n\n\n\u003cp\u003eTotal-ATG14 two-plate assay protocol\u003cbr\u003e\nThe two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a low volume detection plate (either HTRF 384-lv or 96-lv plate) before the addition of HTRF Total-ATG14 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\u003eTotal-ATG14 one-plate assay protocol\u003cbr\u003e\nDetection of Total ATG14 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 4\u003cbr\u003e\nMolecular Modification-Total\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-10,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775251271849,"sku":"64ATG14TPEH","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/es\/products\/revvity-64atg14tpeh","provider":"Iright","version":"1.0","type":"link"}