{"product_id":"revvity-64sh1peh","title":"Revvity, 64SH1PEH, HTRF Human Phospho-SHP1 (Tyr564) Detection Kit, 10,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis cell-based assay is designed to monitor the phosphorylation of SHP1 on Tyr564, which represents a hallmark of its activation.\u003cbr\u003e\nMany cancer cells overexpress checkpoint inhibitor ligands, such as PD-L1. PD-L1 binds its counterpart checkpoint inhibitor receptor PD1, present at the surface of T lymphocytes. In turn, the PD1-PDL1 complex recruits and activates inhibitory effectors, such as SHP1 or SHP2. These two phosphatases, which are phosphorylated on Tyr564 and Tyr542 respectively by the kinase Lck, trigger the dephosphorylation of signaling proteins such as ZAP-70 or SLP-76, involved in the T cell activation pathway. Finally, activated SHP1 and SHP2 participate in T cell inactivation.\u003cbr\u003e\nPreventing the activation of SHP1 and\/or SHP2 by small molecule inhibitors is believed to contribute to restoring the immune response against tumors.\u003cbr\u003e\nHow it works\u003cbr\u003e\nPhospho-SHP1 (Tyr564) assay principle\u003cbr\u003e\nThe Phospho-SHP1 (Tyr564) assay measures SHP1 when phosphorylated at Tyr564. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Phospho-SHP1 (Tyr564) assay uses 2 labeled antibodies, one with a donor fluorophore and the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies, which brings the donor fluorophore into close proximity to the acceptor, thus 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\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-SHP1 (Tyr564) two-plate assay protocol\u003cbr\u003e\nThe two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of Phospho-SHP1 (Tyr564) HTRF detection reagents. This 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\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-SHP1 (Tyr564) one-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated SHP1 (Tyr564) with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nLysis Buffer Compatibility-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\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Infectious Diseases Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-10,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775314579625,"sku":"64SH1PEH","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64sh1peh","provider":"Iright","version":"1.0","type":"link"}