{"product_id":"revvity-alsu-cir1myd-a500","title":"Revvity, ALSU-CIR1MYD-A500, AlphaLISA Human IRAK1 \/ MyD88 Complex Detection Kit, 500 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThe IRAK1\/MYD88 complex forms the initial signaling platform for TLR and IL-1R pathways, serving as the foundation for innate immune activation. Upon receptor engagement, MYD88 oligomerizes and recruits IRAK1 through death domain interactions. MYD88 directly binds IRAK1, bringing it into proximity with IRAK4 for phosphorylation and activation. Oncogenic MYD88 L265P mutations create constitutive IRAK1\/MYD88 complexes that drive malignant B cell survival through NF-κB activation. Disrupting the IRAK1\/MYD88 interaction represents a therapeutic strategy for MYD88-driven lymphomas.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Human IRAK1 \/ MyD88 Complex is a sandwich immunoassay for the quantitative detection of IRAK1 \/ MyD88 complex 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\nAlphaLISA SureFire Ultra protein complex detection assay principle\u003cbr\u003e\nThe AlphaLISA SureFire Ultra complex assay measures cellular protein-protein interactions (PPI).\u003cbr\u003e\nThe assay uses two antibodies which recognize Protein 1 and Protein 2, respectively. AlphaLISA assays require two bead types: Acceptor and Donor Beads. Acceptor Beads are coated with a proprietary CaptSure™ agent to specifically immobilize the first assay antibody, labeled with a CaptSure tag. Donor Beads are coated with streptavidin to capture the second antibody, which is biotinylated. In the presence of the protein complex, 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 PPI 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\u003eAlphaLISA SureFire Ultra complex detection 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 complex AlphaLISA SureFire Ultra detection reagents. This protocol allowing 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\u003eAlphaLISA SureFire Ultra protein complex detection one-plate assay protocol\u003cbr\u003e\nDetection of the protein complex 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 robust AlphaLISA SureFire Ultra quality.\u003cbr\u003e\nAssay sensitivity\u003cbr\u003e\nAssay sensitivity - cell lysate dilution\u003cbr\u003e\nCell lysate was prepared from Karpas 299 cells prepared at 2 x 106 cells\/mL and stimulated with 10 ng\/mL IL-1b for 10 minutes in HBSS + 0.1% BSA. Cells were lysed with the addition of 5X Lysis Buffer for 10 minutes at RT with shaking.\u003cbr\u003e\nLysate was serially diluted in Lysis Buffer and IRAK1\/MyD88 Complex levels were evaluated by AlphaLISA SureFire Ultra. For the detection step, 10 µL of lysate was transferred into a 384-well white OptiPlate, followed by 5 µL of Acceptor mix and incubated for 1 hour at room temperature. 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\nApproximate number of cells per datapoint is indicated. The dotted line represents assay background. The assay can detect IRAK1\/MyD88 Complex down to 500 cells\/datapoint. \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-IRAK1 \/ MyD88\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human\u003cbr\u003e\nTechnology-Alpha\u003cbr\u003e\nTherapeutic Area-Inflammation\u003cbr\u003e\nUnit Size-500 Assay Points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46774629531817,"sku":"ALSU-CIR1MYD-A500","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-cir1myd-a500","provider":"Iright","version":"1.0","type":"link"}