{"product_id":"revvity-alsu-tripk1-b-hv","title":"Revvity, ALSU-TRIPK1-B-HV, AlphaLISA Mouse Total RIPK1 Detection Kit, 100 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nReceptor-interacting serine\/threonine-protein kinase 1 (RIPK1) is a critical regulator of cell death and inflammation. RIPK1 can mediate cell survival, apoptosis, or necroptosis depending on cellular context and post-translational modifications. In its active form, RIPK1 promotes necroptosis by interacting with RIPK3 and MLKL. Dysregulation of RIPK1 activity is implicated in inflammatory and neurodegenerative diseases, as well as in cancer.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Mouse Total RIPK1 Detection Kit is a sandwich immunoassay for the quantitative detection of total RIPK1 in cellular lysates, using Alpha Technology.\u003cbr\u003e\nFormats:\u003c\/p\u003e\n\n\u003cp\u003eThe HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.\u003cbr\u003e\nThe 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.\u003cbr\u003e\nThe 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.\u003cbr\u003e\nThe 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.\u003c\/p\u003e\n\n\u003cp\u003e \u003cbr\u003e\nAlphaLISA SureFire Ultra kits are compatible with:\u003c\/p\u003e\n\n\u003cp\u003eCell and tissue lysates\u003cbr\u003e\nAntibody modulators\u003cbr\u003e\nBiotherapeutic antibodies\u003c\/p\u003e\n\n\u003cp\u003e \u003cbr\u003e\nAlpha SureFire Ultra kits can be used for:\u003c\/p\u003e\n\n\u003cp\u003eCellular kinase assays\u003cbr\u003e\nReceptor activation studies\u003cbr\u003e\nScreening\u003cbr\u003e\nHow it works\u003cbr\u003e\nTotal-AlphaLISA SureFire Ultra assay principle\u003c\/p\u003e\n\n\n    \u003cp\u003eThe Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.\u003c\/p\u003e\n\n\n    \u003cp\u003eThe Total-AlphaLISA SureFire Ultra assay uses two antibodies which recognize two different distal epitopes on the targeted protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure tag. Donor beads are coated with streptavidin to capture one of the detection antibodies, which is biotinylated. In the presence of targeted protein, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of protein present in the sample.\u003c\/p\u003e\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    \u003cp\u003e \u003c\/p\u003e\n\n\n\n\n            \u003cp\u003eTotal-AlphaLISA SureFire Ultra two-plate assay protocol\u003c\/p\u003e\u003cbr\u003e\n        \n\n            \u003cp\u003eThe two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well OptiPlate™ plate before the addition of Total-AlphaLISA SureFire Ultra detection reagents. This protocol permits the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.\u003c\/p\u003e\u003cbr\u003e\n        \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\n            \u003cp\u003eTotal-AlphaLISA SureFire Ultra one-plate assay protocol\u003c\/p\u003e\u003cbr\u003e\n        \n\n            \u003cp\u003eDetection of Total target protein with AlphaLISA SureFire Ultra reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining AlphaLISA SureFire Ultra quality.\u003cbr\u003e\nAssay versatility\u003cbr\u003e\nValidation of mouse RIPK1 total assay in various cell lines\u003cbr\u003e\nHuman and mouse cell lysates were diluted with Lysis Buffer.\u003cbr\u003e\nRIPK1 total levels were evaluated by AlphaLISA SureFire Ultra. For the detection step, 10 µL of cell lysate (approximately 4,000 cells for human cell lines and 8,000 cells for mouse cell lines) were transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor Mix and incubated for 1 hour at RT. Finally, 5 µL of Donor Mix was then added to each well and incubated for 1 hour at RT in the dark. The plate was read on an Envision using standard AlphaLISA settings.\u003cbr\u003e\nAs expected, the Mouse RIPK1 Total AlphaLISA SureFire Ultra assay recognizes RIPK1 Total levels in mouse cell lines only, no signal was detected in human cell lines. \u003cbr\u003e\nApplication-Protein Quantification\u003cbr\u003e\nAutomation Compatible-Yes\u003cbr\u003e\nBrand-AlphaLISA SureFire Ultra\u003cbr\u003e\nDetection Modality-Alpha\u003cbr\u003e\nHost Species-Mouse\u003cbr\u003e\nMolecular Modification-Total\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nShipping Conditions-Shipped in Blue Ice\u003cbr\u003e\nTarget-RIPK1\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Mouse\u003cbr\u003e\nTechnology-Alpha\u003cbr\u003e\nTherapeutic Area-Inflammation\u003cbr\u003e\nUnit Size-100 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46774644277417,"sku":"ALSU-TRIPK1-B-HV","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-tripk1-b-hv","provider":"Iright","version":"1.0","type":"link"}