By Ross D. Brown, P. Joy Ho
A complete selection of vintage and cutting edge methodologies utilized in many laboratories for the research of a number of myeloma. those with no trouble reproducible concepts diversity from the traditional Plasma telephone Labeling Index method to a last bankruptcy on making experience of microarrays, and comprise the entire spectrum of cytogenetic and molecular diagnostic equipment. The protocols stick with the profitable tools in Molecular Medicineв„ў sequence layout, each one providing step by step laboratory directions, an creation outlining the main at the back of the method, lists of the mandatory gear and reagents, and tips about troubleshooting and averting identified pitfalls. those confirmed concepts are perfect for learning the pathogenesis of a number of myeloma and choosing new healing goals.
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Additional resources for Multiple Myeloma: Methods and Protocols
Bone Marrow Culture 1. 0 mL in a sterile syringe or tube containing approx 100 U of preservative-free heparin. 2. RPMI-1640 medium with L-glutamine and HEPES: Store the powdered form at 4°C. 3 using 1 N HCl or 1 N NaOH, and sterilize by filtration. Aseptically decant the medium into sterile bottles and store at 4°C for 1 to 2 wk. The medium should not be used if it becomes opaque, changes color, or acquires floating particles. 3. , 20-mL aliquots for addition to 200-mL bottles of medium to produce a 10% solution).
Immunol. 12, 375–382. Plasma Cell Labeling Index 35 12. Witzig, T. , Kimlinger, T. , Ahmann, G. , Katzmann, J. , and Greipp, P. R. (1996) Detection of myeloma cells in the peripheral blood by flow cytometry. Cytometry 26, 113–120. 13. , and Van Ness, B. (1995) Detection of B-cells clonally related to the tumor population in multiple myeloma and MGUS. Curr. Top. Microbiol. Immunol. 194, 9–16. 14. , and Lichtenstein, A. (1987) Evidence for peripheral blood B lymphocyte but not T lymphocyte involvement in multiple myeloma.
Under inadequate conditions PCs may lose CD138 expression and rapidly enter apoptosis) (21,22). However, if required, storage for longer periods should be performed at lower temperatures (≈2–8°C). In the case of samples aged longer than recommended or any specimens showing evidence of freezing, hemolysis, or clotting, cell viability should be assessed prior to phenotypic studies. The criteria for rejection of sample based on suboptimal conditions that may preclude successful analysis may vary, depending on the specific goals of the immunophenotypic studies.