Amine Reactive Comp-Bead 2 Population Kit

Novus Biologicals | Catalog # NBP3-00496

Novus Biologicals
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Key Product Details

Applications

Flow Cytometry

Kit Type

Kit

Product Summary for Amine Reactive Comp-Bead 2 Population Kit

The Amine Reactive Compensation Beads are designed for labeling amine-reactive dyes (live/dead fixable viability stains) when setting compensation in flow cytometry.
  • Ensure consistent and accurate compensation in any channel
  • Evaluate cell viability of animal cells, bacteria, yeast, and fungi
  • Contains both amine reactive and nonreactive beads for generating positive and negative populations
Concentration: 10^7 particles / mL

Particle size: 7.0 -7.9 micron

Particle material: polystyrene

Product Specifications

Application Notes

Dye and bead concentrations may be further optimized for best results. Centrifugation force and time may need to be increased if needed. Protect the beads from light after exposure to the dye. Use immediately after staining.

Scientific Data Images for Amine Reactive Comp-Bead 2 Population Kit

Amine Reactive Comp-Bead 2 Population [NBP3-00496] - Histogram. Amine Reactive Comp-Beads stained with Biolegend Zombie Green(Tm).
Amine Reactive Comp-Bead 2 Population Kit

Flow Cytometry: Amine Reactive Comp-Bead 2 Population Kit [NBP3-00496]

Flow Cytometry: Amine Reactive Comp-Bead 2 Population Kit [NBP3-00496] - Beads used for flow cytometry compensation calculations. Image from a verified customer review.

Kit Contents for Amine Reactive Comp-Bead 2 Population Kit

  • High Binding Beads - 5 mL
  • Negative Binding Beads - 5 mL

Formulation, Preparation, and Storage

Formulation

0.016 M PBS (pH 7.4)

Preservative

0.02% Sodium Azide

Concentration

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at 4C in the dark. Do not freeze.

Background: Amine Reactive Comp-Bead 2 Population

Compensation is the mathematical correction for the amount of overlap (spillover) of one fluorochrome's emission spectra into another fluorochrome's channel. To calculate this overlap, record single-color stained controls for each fluorochrome in a multicolor panel. This allows the cytometer to visualize the emission of each fluorochrome in each channel, or detector, and subtract the spillover, generating a matrix to apply to multi-color experimental samples. To ensure accurate calculations, single-color controls need to be as bright, or brighter, than the marker on the cell of interest. With increased use of multi-parametric flow cytometry and the large number of fluorochromes available, it is more important than ever to ensure accurate compensation for reliable interpretation of data and reproducible experimental results.

Compensation beads are a highly desirable alternative to traditional cell-based single-color compensation. Positive beads can come pre-loaded with anti-IgG for conjugated antibody capture of mouse, rat, or hamster origin. Uncoated (negative) beads ensure standardization of the autofluorescence in each channel. Beads are preferable to cell-based compensation for several reasons:

1) No precious sample is wasted, and more sample can be used for experimental acquisition.

2) Since 100% of positive beads are able to capture the antibody, the exact experimental fluorochrome can be used regardless of a marker's cellular expression. Dimly expressed markers may not be bright enough for compensation using cells.

3) Reduced autofluorescence of beads allows for more precise calculation of spectral overlap.

Alternate Names

Comp Beads, Compensation Beads, Flow Cytometry Compensation, Live Dead Compensation Beads, Live/Dead Compensation Beads

Additional Amine Reactive Comp-Bead 2 Population Products

Product Documents for Amine Reactive Comp-Bead 2 Population Kit

Certificate of Analysis

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Product Specific Notices for Amine Reactive Comp-Bead 2 Population Kit

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Customer Reviews for Amine Reactive Comp-Bead 2 Population Kit (1)

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  • Amine Reactive Comp-Bead 2 Population Kit
    Name: Shinford Sun
    Verified Customer | Posted 09/16/2024
    We used beads for flow cytometry compensation calculations, and in the image, confidential information has been removed. The different populations are clearly distinguishable following two population comp-bead based compensation.
    Amine Reactive Comp-Bead 2 Population Kit NBP3-00496

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Protocols

View specific protocols for Amine Reactive Comp-Bead 2 Population Kit (NBP3-00496):

1. Allow beads to come to room temperature, then vortex briefly.
2. Add 1 drop (about 50 uL) of the High Binding beads to a 1.5 mL microcentrifuge tube.
3. Wash the beads by adding 0.5 mL of PBS (free of surfactant and blocker) to the microcentrifuge tube, centrifuge at 300 x G for 5 minutes, decant and repeat.
4. Decant and resuspend in 50 uL PBS.
5. Prepare the amine reactive dye according to the manufacturer's instructions.
6. Add 1 - 4 uL of the amine reactive dye to the bead suspension and vortex briefly.
7. Incubate for 30 minutes. Protect tube from light.
8. Add 1 mL of PBS to the same tube and vortex briefly.
9. Centrifuge at 300 x G for 5 minutes, decant and repeat.
10. Resuspend the beads in PBS containing 0.05% BSA with brief vortex.
11. Add a drop (about 50 uL) fo the Negative Binding beads to the labeled High Binding beads.
12. Analyze on the flow cytometer using a live gate around the singlet population in the FSC/SSC dot plot.
13. Create a fluorescent histogram for the appropriate detectors and perform compensation to achieve the desired results.

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FAQs

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