{"product_id":"revvity-64smad4t7peg","title":"Revvity, 64SMAD4T7PEG, HTRF Human and Mouse Phospho-SMAD4 (Thr277) Detection Kit, 500 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis HTRF cell-based assay enables the rapid, quantitative detection of SMAD4 phosphorylated at Threonine 277, as a readout of TGF-ß signaling activity.\u003cbr\u003e\nTGF-ß receptors directly activate the SMAD2\/3 complex by phosphorylation, inducing the association with phospho SMAD4 that then translocates to the nucleus and regulates the gene expression involved in apoptosis, migration, and differentiation, as well as in immune\/inflammatory responses and extracellular matrix remodeling.\u003cbr\u003e\nHTRF assays offer many advantages over other technologies:\u003c\/p\u003e\n\n\u003cp\u003eHomogeneous add-and-read format\u003cbr\u003e\nNo wash steps\u003cbr\u003e\nLow background\u003cbr\u003e\nStraightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates\u003cbr\u003e\nStable signal, providing flexibility in time of readout or size of assays\u003cbr\u003e\nHow it works\u003cbr\u003e\nHTRF phospho SMAD4 Thr277 assay principle\u003cbr\u003e\nThe Phospho-SMAD4 (Thr277) assay measures SMAD4 when phosphorylated at Thr277. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer.\u003cbr\u003e\nThe Phospho-SMAD4 (Thr277) assay uses 2 labeled antibodies: one with a donor fluorophore, the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving the two labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein’s phosphorylation state under a no-wash assay format.\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-SMAD4 (Thr277) 2-plate assay protocol\u003cbr\u003e\nThe 2 plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of the Phospho-SMAD4 (Thr277) HTRF detection reagents.\u003cbr\u003e\nThis protocol enables the cells' viability and confluence to be monitored.\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-SMAD4 (Thr277) one-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated SMAD4 (Thr277) with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required.\u003cbr\u003e\nThis HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling\u003cbr\u003e\nAutomation Compatible-Yes\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nLysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 3 Lysis Buffer 4 Lysis Buffer 5\u003cbr\u003e\nMolecular Modification-Phosphorylation\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nSample Volume-16 µL\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human Mouse\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Cardiovascular Metabolism\/Diabetes NASH\/Fibrosis Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-500 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775267917993,"sku":"64SMAD4T7PEG","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64smad4t7peg","provider":"Iright","version":"1.0","type":"link"}