{"product_id":"revvity-alsu-airak1-a-hv","title":"Revvity, ALSU-AIRAK1-A-HV, AlphaLISA Human IRAK1 Dimer Detection Kit, 100 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nIRAK1 homodimers represent an active signaling state that forms during TLR\/IL-1R pathway activation and contributes to signal amplification. Following IRAK4-mediated phosphorylation, IRAK1 molecules undergo trans-autophosphorylation and form dimeric complexes with enhanced kinase activity. The dimeric form shows increased catalytic efficiency and enhanced ability to recruit TRAF6, amplifying the inflammatory signal. Hyperactive IRAK1 dimerization contributes to excessive inflammation in autoimmune diseases and supports oncogenic signaling in MYD88-mutant cancers. The dimer interface represents a potential therapeutic target for selective IRAK1 inhibition.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Human IRAK1 Dimer is a sandwich immunoassay for the quantitative detection of IRAK1 dimer 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 dimer \/ aggregate detection assay principle\u003cbr\u003e\nThe AlphaLISA SureFire Ultra dimer\/aggregate assay measures the levels of a given protein in a dimeric, multimeric or any other aggregated form.\u003cbr\u003e\nThe assay uses a single antibody clone tagged with both CaptSure™ peptide and biotin, that recognizes exposed epitopes on the multivalent target protein. AlphaLISA assays require two bead types: Acceptor and Donor Beads. Acceptor Beads are coated with a proprietary CaptSure agent to specifically immobilize the antibody labeled with a CaptSure tag. Donor Beads are coated with streptavidin to capture the same specific antibody, which is biotinylated. In the presence of the multivalent 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 multimeric 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\u003eAlphaLISA SureFire Ultra dimer \/ aggregate 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 dimer or aggregate AlphaLISA SureFire Ultra detection reagents. This protocol allows for 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 dimer \/ aggregate detection one-plate assay protocol\u003cbr\u003e\nDetection of the dimer or aggregate 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 Dimer 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 Dimer 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-30 µL\u003cbr\u003e\nShipping Conditions-Shipped in Blue Ice\u003cbr\u003e\nTarget-IRAK1\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human\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":46774629269673,"sku":"ALSU-AIRAK1-A-HV","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-airak1-a-hv","provider":"Iright","version":"1.0","type":"link"}