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A 25 l of mouse plasma was transferred into 1.5 ml centrifuge tube which was spiked with 2.5 l of internal standard (IS) working standard solution (100 g/ml) to get final IS contraction of 250 ng/ml and the solution was vortexed

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It causes an immediate release of almost all the granules (containing serotonin, histamine, chemokines, cytokines, enzymes, heparin, and prostaglandins), which trigger itching, swelling, mucous, and anaphylaxis.[ref] A delayed response by mast cells comes from the formation of other mediators, such as prostaglandin D2