Platelet surface β1–4 galactose exposure using FITC-conjugated R. communis agglutinin I (RCA-I, Vector Labs) and FITC-conjugated E. cristagalli lectin (ECL, Vector Labs), and Mpl expression were determined by flow cytometry.20 (link) To determine Mpl internalization platelets were incubated with 50 ng ml−1 TPO for 10 min at 37 °C or not (rest), then all samples were incubated with a rabbit antibody directed against the extracellular domain of Mpl (provided by Dr. Wei Tong, UPenn) or control rabbit IgG, followed by Alexa Fluor 488-labeled goat anti-rabbit IgG antibody, and analyzed by flow cytometry.
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Platelet Surface Glycan and Receptor Analysis
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Agglutinin
Alexa fluor 488
Anti igg
Antibody
Fitc
Flow cytometry
Galactose
Goat
Lectin
Platelets
Rabbit
Vector
Corresponding Organization :
Other organizations : Brigham and Women's Hospital, Harvard University, Boston Children's Hospital
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Variable analysis
independent variables
- Treatment with 50 ng ml^-1 TPO for 10 min at 37 °C
- Platelet surface β1–4 galactose exposure using FITC-conjugated Ricinus communis agglutinin I (RCA-I) and FITC-conjugated Erythrina cristagalli lectin (ECL)
- Mpl expression
- Mpl internalization
- Untreated platelets (rest)
- Rabbit antibody directed against the extracellular domain of Mpl
- Control rabbit IgG
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