{"product_id":"revvity-64n62peg","title":"Revvity, 64N62PEG, HTRF Human and Mouse Total p62 (SQSTM1) Detection Kit, 500 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis HTRF cell-based assay conveniently and accurately monitors the expression level of p62\/SQSTM1 proteins, both phosphorylated and unphosphorylated. It uses the same buffers as our Phospho-p62\/SQSTM1 kit, and enables the analysis of phosphorylated and total proteins from a single sample as a pivotal readout of the autophagy pathway. Autophagy is a cellular mechanism which enables the clearance of abnormal proteins or organelles, and plays a role in cancer, neurodegenerative diseases, as well as cardiovascular and infectious diseases.\u003cbr\u003e\np62\/SQSTM1 is a ubiquitin-binding protein and functions as a bridge between polyubiquitinated cargo protein and autophagosomes. p62 interacts directly with both the cargo protein and a protein called LC3 which is involved in the autophagosome formation. Finally, the activated autophagy pathway promotes the degradation of p62-associated ubiquitinated protein cargoes.\u003cbr\u003e\nHow it works\u003cbr\u003e\nTotal p62\/SQSTM1 assay principle\u003cbr\u003e\nThe Total-p62\/SQSTM1 assay quantifies the expression level of p62 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total-p62\/SQSTM1 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 p62\/SQSTM1 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\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003eTotal-p62\/SQSTM1 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-p62\/SQSTM1 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-p62\/SQSTM1 (Ser403) one-plate assay protocol\u003cbr\u003e\nDetection of Total p62\/SQSTM1 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 Lysis Buffer 5\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 Mouse\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Cardiovascular Infectious Diseases Neuroscience Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-500 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775297704105,"sku":"64N62PEG","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64n62peg","provider":"Iright","version":"1.0","type":"link"}