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Platelet Serotonin ELISA Protocol for Laboratory Analysis

This protocol provides step-by-step instructions for the purification and analysis of platelet serotonin using an ELISA method. It includes details on centrifugation, sample storage, and specimen handling techniques.

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Platelet Serotonin ELISA Protocol for Laboratory Analysis

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  1. Platelet Serotonin Laboratory Protocol Centrifuge 10 minutes at 200 RCF(xg) at ambient temperature 4 mL EDTA tube • Remove 400 uL of plasma • Add to 15 mL tube for platelet purification • Add 1.5 mL physiological saline to platelet-rich plasma (PRP) Centrifuge10 minutes at 4,500 RCF(xg) at 4oC & discard supernatant • Add to 300 uL distilled water • Vortex pellet until the platelets are mixed & transfer 300 uL to 1 aliquot ELISA Transfer to -80oC freezer for storage Immuno Biological Laboratories (2009). Serotonin ELISA IB89546. [Laboratory instructions]. IBL-America

  2. Factors Affecting Biological Specimens Specimen Storage Processing Phlebotomy Specimen Transport • Storage temperature • Duration of storage • Freeze-thaw cycles • Instrument calibration & controls • Time between collection & centrifugation • Vortex technique • Centrifugation • Type • Calibrated • Temperature • Speed • Duration • Automated decapping • Specimen separated into aliquots • Aliquot labeling • In Vivo Haemolysis due to patient condition • Proper labeling • Mode of collection • Needle gauge • Position of arm • Location of venipuncture • Antiseptic used • Tourniquet time • Syringe transfer • Specimen tube • Type • Under/over filled • Order of draw • Under/over mixing • Origin of specimen • Vertical or horizontal transportation of tube • Pneumatic tube transport • Courier transport • Transportation duration • Centrifugation prior to transfer • Transfer temperature

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