{"product_id":"revvity-64lc3b2peh","title":"Revvity, 64LC3B2PEH, HTRF Human and Mouse LC3B II Detection Kit, 10,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nLC3 is a mammalian homolog of the yeast ATG8 protein and is distributed throughout mammalian tissues and cultured cells. LC3 is a key component of the autophagy process - a degradative program that maintains cellular homeostasis.\u003cbr\u003e\nDuring autophagy, cytoplasmic components such as cytosolic proteins and organelles are engulfed by autophagosomes Concurrently, a cytosolic form of LC3 (LC3-I) is conjugated to phosphatidylethanolamine to form LC3-phosphatidylethanolamine conjugate (LC3-II). This newly formed conjugate is recruited to autophagosomal membranes. The subsequent fusion of autophagosomes with lysosomes results in the formation of autolysosomes, and intra-autophagosomal components are degraded by lysosomal hydrolases. At the same time, LC3-II within the autolysosomal lumen is degraded.\u003cbr\u003e\nThe LC3-II level has been found to be proportional to autophagosome abundance making LC3-II the most popular marker of autophagy.\u003cbr\u003e\nHow it works\u003cbr\u003e\nLC3B-II assay principle\u003cbr\u003e\nThe HTRF LC3B-II assay quantifies the expression level of LC3B-II in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The LC3B-II 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 LC3B-II 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\u003eLC3B-II two-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 LC3B-II 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\n\n\n\n\u003cp\u003eLC3B-II one-plate assay protocol\u003cbr\u003e\nDetection of LC3B-II 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-Protein Quantification\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nSample Volume-16 µL\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Biomarkers\u003cbr\u003e\nTarget Species-Human Mouse\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nUnit Size-10,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775266214057,"sku":"64LC3B2PEH","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64lc3b2peh","provider":"Iright","version":"1.0","type":"link"}