{"product_id":"revvity-alsu-tmsh3-a-hv","title":"Revvity, ALSU-TMSH3-A-HV, AlphaLISA Human Total MSH3 Detection Kit, 100 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nMutS Homolog 3 (MSH3) is a DNA mismatch repair protein that forms the MutSβ complex with MSH2 to recognize and repair insertion-deletion loops (IDLs) and larger mismatches in DNA. MSH3 is particularly important for repairing slipped-strand intermediates that arise during DNA replication in repetitive sequences, including microsatellites. The MutSβ complex initiates repair by recruiting MLH1-PMS2 (MutLα) and other downstream factors to excise and resynthesize the error-containing DNA strand. Deficiency in MSH3 leads to microsatellite instability (MSI) and increased mutation rates, particularly in repetitive sequences. MSH3 mutations are associated with certain hereditary cancer syndromes and contribute to tumorigenesis through genomic instability. In cancer therapy, MSH3 status influences response to DNA-damaging agents and immunotherapy, as MSI tumors often exhibit enhanced immunogenicity.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Human Total MSH3 is a sandwich immunoassay for the quantitative detection of total MSH3 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\nEndogenous MSH3 expression 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 100 µL of Lysis Buffer. Suspension cells were washed with HBSS and lysed with Lysis Buffer at 4 x 106 cells\/mL.\u003cbr\u003e\nMSH3 levels were evaluated by AlphaLISA SureFire Ultra. For the detection step, 10 µL of cell lysate (4,000 adherent cells or 40,000 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\nEndogenous levels of MSH3 were detected in various cell types. \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-30 µL\u003cbr\u003e\nShipping Conditions-Shipped in Blue Ice\u003cbr\u003e\nTarget-MSH3\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human\u003cbr\u003e\nTechnology-Alpha\u003cbr\u003e\nTherapeutic Area-Neuroscience Oncology\u003cbr\u003e\nUnit Size-100 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46774645129385,"sku":"ALSU-TMSH3-A-HV","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-tmsh3-a-hv","provider":"Iright","version":"1.0","type":"link"}