{"product_id":"revvity-alsu-tmcl1-a50k","title":"Revvity, ALSU-TMCL1-A50K, AlphaLISA Human and Mouse Total MCL-1 Detection Kit, 50,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nMyeloid Cell Leukemia 1 (MCL-1) is an anti-apoptotic protein belonging to the BCL-2 family that prevents mitochondrial outer membrane permeabilization and cytochrome c release. MCL-1 localizes to the mitochondrial outer membrane where it sequesters pro-apoptotic proteins such as BAK, BAX, and BH3-only proteins, thereby blocking apoptosis initiation. MCL-1 expression is tightly regulated at transcriptional, post-transcriptional, and post-translational levels, with rapid turnover under normal conditions. Overexpression of MCL-1 is frequently observed in hematologic malignancies and solid tumors, where it confers resistance to chemotherapy and targeted therapies. MCL-1 dependency varies among cancer types, making it an attractive target for selective cancer therapy. BH3 mimetics and MCL-1-specific inhibitors are being developed to overcome apoptotic resistance in MCL-1-dependent cancers.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Human and Mouse Total MCL-1 is a sandwich immunoassay for the quantitative detection of total MCL-1 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\u003eAlphaLISA 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\u003eAlphaLISA SureFire Ultra kits can be used for:\u003c\/p\u003e\n\n\u003cp\u003eCellular kinase assays\u003cbr\u003e\nReceptor activation studies\u003cbr\u003e\nHigh-throughput screening for preclinical studies\u003cbr\u003e\nHow it works\u003cbr\u003e\nTotal-AlphaLISA SureFire Ultra assay principle\u003cbr\u003e\nThe Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.\u003cbr\u003e\nThe 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  \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\u003cp\u003eTotal-AlphaLISA SureFire Ultra two-plate assay protocol\u003cbr\u003e\nThe 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\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-AlphaLISA SureFire Ultra one-plate assay protocol\u003cbr\u003e\nDetection 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\nExpression of MCL-1 in various cell lines\u003cbr\u003e\nAdherent cells were seeded at 40,000 cells\/well in a 96-well culture plate in complete medium and incubated overnight at 37°C, 5% CO2. Cells were lysed with 200 µL of Lysis Buffer.\u003cbr\u003e\nSuspension cells were seeded at 400,000 cells\/well in a 96-well culture plate in HBSS + 0.1% BSA, cells were spun down and lysed with 200 µL of Lysis Buffer. MCL-1 Total levels were evaluated by AlphaLISA SureFire Ultra. For the detection step, 10 µL of cell lysate (approximately 2,000 adherent cells or 20,000 suspension cells) was 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\nMCL-1 is broadly expressed in various cell lines. High MCL-1 expression in observed in Raji and EL-4 cells while lower levels were detected in Hep G2 and K-562 cells. \u003cbr\u003e\nApplication-Cell Signaling\u003cbr\u003e\nAutomation Compatible-Yes\u003cbr\u003e\nBrand-AlphaLISA SureFire Ultra\u003cbr\u003e\nDetection Modality-Alpha\u003cbr\u003e\nProtocol Time-2h at RT\u003cbr\u003e\nSample Volume-10 µL\u003cbr\u003e\nShipping Conditions-Shipped in Blue Ice\u003cbr\u003e\nTarget-MCL-1\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human Mouse\u003cbr\u003e\nTechnology-Alpha\u003cbr\u003e\nTherapeutic Area-Oncology\u003cbr\u003e\nUnit Size-50,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46774636380329,"sku":"ALSU-TMCL1-A50K","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-tmcl1-a50k","provider":"Iright","version":"1.0","type":"link"}