BD™ SpectraComp™ Unmixing and Compensation Particles
Preparation And Storage
Recommended Assay Procedures
The BD™ SpectraComp™ Unmixing and Compensation Particles have been tested with mouse, rat, and hamster Ig antibodies conjugated to various fluorochromes. See the specific instructions below on the use of the BD™ SpectraComp™ Unmixing and Compensation Particles.
1. Vortex BD™ SpectraComp™ Unmixing and Compensation Particles thoroughly before use.
2. Add 1 drop (approximately 50 µl) of BD™ SpectraComp™ Unmixing and Compensation Particles into the bottom of 12 × 75 mm tubes or 96-well plate wells for each fluorochrome used in the experiment. It is recommended to include one tube for unstained control.
3. Add antibody reagent to the mixture and vortex. Incubate at room temperature for 15-30 minutes, protected from light.
Notes:
• It is recommended to predetermine the appropriate amount of antibody reagent that works best for the application.
• Treat the BD™ SpectraComp™ Unmixing and Compensation Particles with the same protocol used for the cell sample, eg, if the testing protocol uses BD Horizon™ Brilliant Stain Buffer or permeabilized and fixed cells, the particles should also be treated with the reagents and protocol.
4. Add 2 mL of staining buffer [eg, BD Pharmingen™ Stain (FBS), Cat. No. 554656 or BD Pharmingen™ Stain (BSA), Cat. No. 554657] to the tubes or required volume to the plate wells.
5. Wash the particles by centrifuging the tubes or plate for 5 minutes at 500 × g and immediately aspirate the supernatants to minimize particle loss, being careful not to disturb the pellet. Wash two times by repeating steps 4 and 5 leaving approximately 50 µL of supernatant in the tube after each aspiration.
Note:
• For best signal-to-noise results, use a vacuum aspirator and aspirate off the supernatant using a fine pipette tip.
6. Resuspend the pellet by vortexing and by then adding 0.3-0.5 mL staining buffer.
7. Acquire the BD™ SpectraComp™ Unmixing and Compensation Particles using the same Forward and Side Light Scatter parameter (FSC-A and SSC-A) instrument settings used for analyzing cells. It is recommended to acquire the data immediately after staining.
8. Gate on the singlet bead population based on FSC (forward-light scatter) and SSC (side-light scatter) characteristics.
9. For compensation or spectral unmixing use the appropriate protocol for your cytometer system.
10. Proceed to acquire the cell samples for the experiment.
It is recommended to test the BD™ SpectraComp™ Unmixing and Compensation Particles to confirm accuracy of compensation or spectral unmixing.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- This product was developed and manufactured by Slingshot Biosciences, Inc., Emeryville, CA.
Companion Products
The BD™ SpectraComp™ Unmixing and Compensation Particles are microparticles used to help generate fluorochrome profiles for spectral unmixing and compensation in multicolor flow cytometry. The product contains both a positive and negative particle in the same dropper bottle. The positive particle can bind both kappa and lambda light chain-bearing immunoglobulins raised in mouse, rat, and hamster hosts. When mixed with fluorochrome-conjugated antibodies, the BD™ SpectraComp™ Unmixing and Compensation Particles provide distinct positive and negative populations that can be used to set up instruments so that spectral overlap can be corrected.
The BD™ SpectraComp™ Unmixing and Compensation Particles are compatible with legacy base and tandem fluorochromes including BD Horizon Brilliant™ dyes as well as BD Horizon RealYellow™ and BD Horizon RealBlue™ fluorochromes. In addition, they are compatible with commonly used flow cytometry buffers, such as, BD Horizon™ Brilliant Stain Buffer, BD FACS™ Lysing Solution, BD Pharm Lyse™ Lysing Buffer, BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit, BD Cytofix™ Fixation Buffer, BD Pharmingen™ Transcription Factor Buffer Set, and BD Phosflow™ Perm Buffer III.
No Citations Are Available for this Product
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.