{"product_id":"revvity-alsu-phish1-a50k","title":"Revvity, ALSU-PHISH1-A50K, AlphaLISA Human Phospho-Histone H1 (Thr17) Detection Kit, 50,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nHistone H1 is a linker histone that binds to nucleosomes and linker DNA to stabilize higher-order chromatin structure and regulate DNA accessibility. H1 binds to the exterior of the nucleosome, compacting chromatin into 30-nm fibers and higher-order structures. Mammals express multiple H1 variants with distinct expression patterns and regulatory functions. H1 binding is dynamically regulated through post-translational modifications that modulate chromatin compaction and gene expression. Altered H1 expression and mutations are found in various cancers and developmental disorders, affecting genome stability and transcriptional regulation.\u003cbr\u003e\nThe AlphaLISA SureFire Ultra Human Phospho-Histone H1 (Thr17) is a sandwich immunoassay for the quantitative detection of phospho-Histone H1 (Thr17) 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\nPhospho-AlphaLISA SureFire Ultra assay principle\u003cbr\u003e\nThe Phospho-AlphaLISA SureFire Ultra assay measures a protein target when phosphorylated at a specific residue.\u003cbr\u003e\nThe assay uses two antibodies which recognize the phospho epitope and a distal epitope 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 phosphorylated 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 phosphoprotein 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\u003ePhospho-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 Phospho-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\u003ePhospho-AlphaLISA SureFire Ultra one-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated 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 sensitivity\u003cbr\u003e\nAssay sensitivity - cell lysate dilution\u003cbr\u003e\nCell lysate was prepared from HEK293 cells cultured to confluence in a T175 flask and lysed in 40 mL of Lysis Buffer B for 10 minutes at RT with shaking.\u003cbr\u003e\nLysate was serially diluted in Lysis Buffer and Histone H1 Phospho (Thr17) levels were evaluated by AlphaLISA SureFire Ultra. For the detection step, 10 µL of cell 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\nApproximate number of cells per datapoint is indicated. The dotted line represents assay background. The assay can detect Histone H1 (Thr17) in HEK293 cells in less than 200 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-Histone H1\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human\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":46774624288937,"sku":"ALSU-PHISH1-A50K","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-alsu-phish1-a50k","provider":"Iright","version":"1.0","type":"link"}