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Boster offers custom polyclonal antibody production for researchers who use non-mammalian models such as Zebrafish, Drophila, C. elegans and Yeast at $600. Contact us for a free consultation.
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Protocols, optimization tips,troubleshooting guides, and more for flow cytometry.
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Note: This information is to serve as a guide as individual investigators may need to optimize protocols for their particular cell type.
Cells for flow cytometry analysis are usually derived from four main sources:
Regardless of the source, the final cell preparation should be:
PBMCs isolated from whole blood through Ficoll gradient centrifugation or RBC lysed whole blood or non-adherent cultured cells are readily available for flow cytometric analysis. Adherent cultured cells or cells present in the solid organs should be first made into a single cell suspension before flow analysis by using enzymatic digestion or mechanical dissociation of the tissue, respectively. Mechanical filtration should be followed to avoid unwanted instrument clogs or lower quality flow data. Use the following sample preparation protocols based on your appropriate starting materials:
Key Reagents – PBS, staining buffer
Key Reagents – PBS, staining buffer, 0.25% trypsin
Key reagents – PBS, staining buffer, culture medium with 10% FBS
Key reagents – PBS, staining buffer, suitable gradient medium like Ficoll or Histopaque
Key reagents – PBS, staining buffer (see appendix), RBC lysis buffer [check Appendix for recipe or use a commercially available buffer]