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ISH/FISH
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Preparation Protocol for Buffy Coat from Whole Blood
GUIDELINE
The buffy coat is a fraction of whole blood that is rich in white blood cells (WBCs) and is named after its "buff-like" color. Preparation of a buffy coat from whole blood samples precludes many downstream applications. During processing, whole blood is centrifuged to form distinct blood fractions.
METHODS
- Add an equal volume of recommended medium to whole blood and mix gently.
- Centrifuge at 800 x g for 10 minutes at room temperature (15-25°C) with the brake off.
- Remove the concentrated leukocyte band (this is the buffy coat), plus a small portion of the plasma and concentrated red blood cells (RBCs). The target is to concentrate the leukocytes approximately 5-fold while maintaining an equivalent ratio of leukocytes to RBCs (e.g. collect 2 mL of buffy coat when starting with 10 mL of whole blood).
Creative Bioarray Relevant Recommendations
- Creative Bioarray offers different ranges of human blood samples, such as whole blood, blood serum, plasma, prothrombins, and coagulation factors. In addition, we provide the pharmaceutical and biotech industry with sample collection and analysis services to researchers worldwide.
- We also provide three types of Buffy Coat Isolation Kit, including the Human HLA Buffy Coat CD56 Isolation Kit, Human CD14 Cell Isolation Kit (Buffy Coat), and Human CD56 Cell Isolation Kit (Buffy Coat). These kits are fast, efficient, and easy to use. No column is required for use and they have a high purity of 99% and high recovery.
NOTES
- Invert the media and whole blood to mix gently.
- Remove the sample from the centrifuge carefully, so as not to disturb the distinct blood fractions.
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