I have platelet pellet in PBS in my lab, but I am not sure how to proceed with platelet adhesion assay using platelet pellet with minimal amount of PBS. Most of the protocol show platelet adhesion assay using fresh human platelet rich plasma (PRP) instead.
Platelet adhesion assay using a platelet pellet:
Materials needed:
- Platelet pellet (obtained by centrifuging PRP at 1000xg for 10 minutes)
- PBS (phosphate-buffered saline)
- Fibrinogen (human or rabbit)
- Thrombin (human or recombinant)
- Prostaglandin E1 (PGE1) (optional)
- Aggregometry cuvettes (e.g., Chronolog)
- Aggregometer (e.g., Chronolog)
- Microcentrifuge tubes (1.5 and 0.5 mL)
- Pipettes (100 μL and 1 mL)
Protocol:
Step 1: Preparation of Platelet Pellet Suspension
Step 2: Preparation of Fibrinogen-Coated Cuvettes
Step 3: Platelet Adhesion Assay
Step 4: Data Analysis
Note: You can modify the concentration of fibrinogen, thrombin, and PGE1 according to your experimental requirements. Additionally, you can include appropriate controls (e.g., negative control without thrombin or positive control with collagen plus thrombin) to validate the results. Always follow proper safety protocols when working with human blood products and biohazardous materials.
All the best
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