Preparation And Storage
• Store the reagent at 2–8 °C.• Reagent in unopened vials is stable until the expiration date shown on the label when stored as directed. Do not use after the expiration date.• Use reagent within 12 months of opening the vial or until the expiration date, whichever comes first, when stored as directed.• Do not freeze the reagent or expose it to direct light during storage or incubation with cells. Keep the reagent vial dry.
Recommended Assay Procedures
1. Add the appropriate volume of CD27 (O323) fluorochrome-conjugated monoclonal antibody to 100 μL of whole blood in a 12 × 75-mm capped polystyrene test tube.
NOTE If using BD Horizon™ Brilliant Stain Buffer, add 50 μL of buffer to the empty tube, then add the fluorochrome-conjugated antibodies, and lastly add 100 μL of whole blood. See the BD Horizon™ Brilliant Stain Buffer IFU.
Fluorochrome Volume per test (μL)
BV510 5
2. Vortex gently and incubate for 15–30 minutes at room temperature (20–25 °C), protected from light.
3. Add 2 mL of 1X BD FACS™ Lysing Solution to each tube.
4. Vortex the tube 3–5 seconds at low speed and incubate for 10 minutes at room temperature, protected from light.
5. Centrifuge at 300g for 5 minutes.
6. Aspirate the supernatant without disturbing the cell pellet.
7. Add 2 mL of buffer to each tube according to the fluorochrome you are using.
8. Vortex gently.
9. Centrifuge at 200g for 5 minutes.
10. Aspirate the supernatant without disturbing the cell pellet.
11. Add 0.5 mL of wash buffer to each tube and acquire the samples immediately.
Optional: Instead of adding buffer, fix the stained sample as described in the following section.
Fixing the Stained Sample (optional)
1. Add 0.5 mL of fixative solution.
2. Vortex gently.
3. Incubate for 60 minutes at 2–8 °C, protected from light.
4. Centrifuge at 300g for 5 minutes.
5. Aspirate the supernatant without disturbing the cell pellet.
6. Add 0.5 mL of wash buffer to each tube.
7. Vortex gently.
Store fixed samples at 2–8 °C, protected from light, until acquisition.
In the absence of BD Horizon™ Brilliant Stain Buffer, we recommend acquiring the samples within 24 hours of staining.
If you are staining cells using more than one BD Horizon Brilliant™ Violet reagent in the presence of BD Horizon™ Brilliant Stain Buffer, we recommend acquiring the sample within 6 hours of fixation.
CAUTION Prolonged exposure of cells to paraformaldehyde can lead to increased autofluorescence in the violet channels. Therefore, cells are centrifuged and resuspended in buffer without paraformaldehyde after 1 hour of fixation.
Development References (13)
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Centers for Disease Control and Prevention. 2007 Guideline for Isolation Precautions: Preventing Transmission of Infectious Agents in Healthcare Settings. Available: https://www.cdc.gov/infectioncontrol/guidelines/isolation/index.html March 12, 2019. View Reference
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Clinical and Laboratory Standards Institute. Collection of Diagnostic Venous Blood Specimens, 7th ed. In: CLSI. CLSI, ed. CLSI document GP41-A7. Wayne, PA: Clinical and Laboratory Standards Institute; 2017:1-85. View Reference
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Clinical and Laboratory Standards Institute. Protection of Laboratory Workers from Occupationally Acquired Infections. In: CLSI. CLSI, ed. CLSI document M29-A4. Wayne, PA: Clinical and Laboratory Standards Institute; 2014:1-133. View Reference
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Gaiti F, Chaligne R, Gu H, et al. Epigenetic evolution and lineage histories of chronic lymphocytic leukaemia.. Nature. 2019; 569(7757):576-580. (Methodology). View Reference
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Jackson AL, Warner NL. Preparation, staining, and analysis by flow cytometry of peripheral blood leukocytes. In: Rose NR, Friedman H, Fahey JL, ed. Manual of Clincial Laboratory Immunology, Third Edition. Washington DC: American Society for Microbiology; 1986:226-235.
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Kroll MH. Evaluating interference caused by lipemia.. Clin Chem. 2004; 50(11):1968-9. (Methodology). View Reference
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Ledergor G, Weiner A, Zada M, et al. Single cell dissection of plasma cell heterogeneity in symptomatic and asymptomatic myeloma.. Nat Med. 2018; 24(12):1867-1876. (Methodology). View Reference
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Mizuta S, Yamane N, Mononobe S, et al. VS38 staining contributes to a novel gating strategy in flow cytometry for small B cell lymphoma, especially in lymphoplasmacytic lymphoma/Waldenström macroglobulinemia.. Cytometry B Clin Cytom. 2022; 102(1):50-61. (Methodology). View Reference
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Nikolac N. Lipemia: causes, interference mechanisms, detection and management.. Biochem Med (Zagreb). 2014; 24(1):57-67. (Methodology). View Reference
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Rodrigues-Santos P, López-Sejas N, Almeida JS, et al. Effect of Age on NK Cell Compartment in Chronic Myeloid Leukemia Patients Treated With Tyrosine Kinase Inhibitors.. Front Immunol. 2018; 9:2587. (Methodology). View Reference
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Rothe G, Schmitz G. Consensus protocol for the flow cytometric immunophenotyping of hematopoietic malignancies. Leukemia. 1996; 10:877-895. (Methodology).
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Stelzer GT, Marti G, Hurley A, McCoy P Jr,Lovett EJ, Schwartz A. US-Canadian Consensusrecommendations on the immunophenotypicanalysis of hematologic neoplasia by flowcytometry: standardization and validation oflaboratory procedures. Cytometry. 1997; 30:214-230. (Methodology).
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Stetler-Stevenson M, Ahmad E, Barnett D et al. Clinical Flow Cytometric Analysis of Neoplastic Hematolymphoid Cells; Approved Guideline- Second Edition CLSI document H43-A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2005.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Documents are subject to revision without notice. Please verify you have the correct revision of the document, and always refer back to BD's eIFU website for the latest and most up to date information.