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 CD138 (MI15) 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)
FITC 20
PE 20
PerCP-Cy5.5 20
APC 5
BV421 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 to 3 mL of wash buffer to each tube.
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 wash 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.
The CD138 antibody, clone MI15, is derived from the hybridization of SP2/0 mouse myeloma cells with spleen cells isolated from BALB/c mice immunized with human cell line 8266.
The CD138 antibody recognizes syndecan-1, a member of the syndecan family of transmembrane heparin sulfate proteoglycans whose main functions are to modulate ligand-dependent activation of primary signaling receptors at the cell surface. Syndecan-1 exhibits molecular polymorphism| its core protein is a single-chain molecule of 70 to 75 kilodaltons (kDa), and its isoforms vary in molecular weight up to approximately 400 kDa.
Development References (21)
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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.
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.