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ArC™ Amine Reactive Compensation Bead Kit (for use with LIVE/DEAD™ Fixable dead cell stain kits), 1 Kit - FAQs

ArC™ Amine Reactive Compensation Bead Kit (for use with LIVE/DEAD™ Fixable dead cell stain kits), 1 Kit - FAQs

View additional product information for ArC™ Amine Reactive Compensation Bead Kit (for use with LIVE/DEAD™ Fixable dead cell stain kits) - FAQs (A10346, A10628)

11 product FAQs found

Are the vials of ArC Amine Reactive Compensation Bead Kit (for use with LIVE/DEAD Fixable dead cell stain kits) (Cat. No. A10346, A10648) compatible with Vaporized Hydrogen Peroxide (VHP) sterilization?

While the polypropylene cap is compatible with Vaporized Hydrogen Peroxide (VHP) sterilization, we have not tested the compatibility of the Low-Density Polyethylene (LDPE) / Linear Low-Density Polyethylene (LLDPE) vials with VHP sterilization.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What is the stability for the ArC Amine Reactive Compensation Bead Kit (for use with LIVE/DEAD Fixable dead cell stain kits)?

When stored as directed (2-8 degrees C), this kit is stable for at least 1 year. Please do not freeze the kit. Please visit the user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/arc_amine_reactive_comp_bead_kit_man.pdf) for further technical details.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

My ArC Amine Reactive Compensation beads are excessively clumped and cannot be resuspended by vortexing or sonication. What may have caused this?

This can occur if the solution of beads was ever frozen. The formation of ice excludes the beads and compresses the beads together. Once frozen, the beads cannot be resuspended into solution and are no longer usable.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

For the ArC Amine Reactive Compensation Bead Kit, why must I use freshly prepared amine-reactive dye?

This is because succinimidyl ester (NHS ester) groups are very labile to water; they dissociate readily from the dye structure if held in solvents that are contaminated with water, as is possible from condensation or absorbing water from the atmosphere.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Besides the Live/Dead Fixable Dead Cell stain, may I conjugate other amine-reactive dyes to the beads in the ArC Amine Reactive Compensation Bead Kit?

Yes. The succinimidyl ester (NHS ester) and isothiocyanate (e.g., FITC, TRITC) dye derivatives may be attached to the ArC beads, but you would have to optimize the amount of reactive dye per ArC beads.

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I am performing flow cytometry. My compensation matrix has values over 100. That's not possible is it?

It is possible. But if you have a lot of values over 100, you probably need to look at the voltages that you are using for your data collection.

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I’ve been told that I need to compensate my data for flow cytometry analysis. What does that mean?

By running single-color controls, it is possible to remove signal of one fluorophore that spills over into the collection channel for another fluorophore.

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What kind of controls do I need for flow cytometry?

For compensation, you need to prepare a singly stained sample (or compensation beads) for each color parameter that you are using. In addition, we recommend that you use FMO (flow minus one) controls. These are controls in which you label cells or beads with every color in your panel, omitting one. Make one FMO control for each color. These controls are important for helping you properly set gates on your data.

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Do I need compensation controls for non-antibody reagents?

Yes, you should have a compensation control for each color in your assay. For amine-reactive stains such as our LIVE/DEAD Fixable Dead Cell Stain Kits, you can label beads to make compensation controls using our ArC Amine Reactive Compensation Bead Kit (Cat. No. A10346). For other stains, such as cell cycle stains, you need to use labeled cells for your compensation control.

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Why should I worry about compensation in flow cytometry analysis?

In a perfect world, the fluorescence emission profile for each individual fluorophore would be a very intense, narrow peak, well separated from all other emission peaks. In reality, organic dyes and fluorescent proteins have broad emission peaks, and compensation must be employed (during or after data acquisition) to correctly assign fluorescence signal to each fluorophore. Compensation is important because it removes fluorescent signal from overlapping spectra so you know that the signal you see is only the signal from the fluorophore of interest.

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How do I make compensation controls for antibodies?

All that you need for compensation controls is an unstained sample of your cells (or negative control beads) and samples with each of your fluorophores - one per tube. You want the single-stained controls to be as bright as you expect your brightest sample to be. Antibodies can be bound to beads instead of cells using our AbC Total Antibody Compensation Bead Kit (Cat. Nos. A10513 and A10497).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.