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HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells
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JoVE Journal Chemistry
HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells

HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells

DOI:
10.3791/54124-v

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11:29 min

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July 20, 2016

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,
1Department of Medical Sciences, Human Genetics Foundation (HuGeF),University of Turin

Chapters

  • 00:05Title
  • 00:59Isolation of Leukemic Lymphocytes from Blood Samples of CCL Patients
  • 02:09Purification of Leukemic B Cells by Negative Isolation
  • 03:27Program the HPLC Method
  • 05:07Pretreatment with the Inhibitors and Incubation with the Substrate (AMP)
  • 06:16Samples Preparation for HPLC and HPLC Measurements of Purines
  • 08:14Results: Quantification of Purine Compounds in Freshly Purified CLL Cells from a Representative CD73+ Patient
  • 09:36Conclusion

Summary

Automatic Translation

Automatic Translation

The protocol described here represents an easy and reproducible method that employs reverse phase high-performance liquid chromatography (RP-HPLC) to measure purine metabolism on chronic lymphocytic leukemia (CLL) cells cultured under different conditions.

Tags

Keywords: HPLCNucleotide MetabolismNucleoside MetabolismChronic Lymphocytic LeukemiaCLL CellsPurine MetabolismAdenosineExtracellular EnvironmentCell CultureDensity Gradient CentrifugationPBMCMagnetic BeadsCell Purification

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