Invitrogen™ UltraComp eBeads™ Compensation Beads

Product Code.	Quantity	unitSize
15536296	25 tests	25 tests
15385356	100 tests	100 tests
16881320	100 tests	Each

Product Code. 15536296 Supplier Invitrogen™ Supplier No. 01222241

Description

Designed for use in compensation with all fluorochromes excited by ultraviolet, violet, blue, green, yellow-green, and red lasers. UltraComp eBeads™ react with antibodies of mouse, rat, and hamster origin, and are immunoglobulin light chain-independent. 1 drop (50μL)/test.

• Each drop of beads contains both a positive population that will capture any mouse, rat or hamster antibody and a negative population that will not react with antibody
• This bimodal distribution can be used for single-color compensation controls in multicolor flow cytometry experiments
• Compatible with all fluorochromes excited by an ultraviolet (355nm) or violet (405nm) laser
• Should be used with standard staining buffers which contain PBS or HBSS, protein such as BSA or FBS, and sodium azide. No other additives should be used.

Specifications

• For Use With (Equipment): Flow Cytometer
• No. of Tests: 25 tests
• Quantity: 25 tests
• Product Line: eBeads
• Product Type: Compensation Beads

Frequently Asked Questions (FAQs)

How long are UltraComp eBeads Compensation Beads (Cat. No. 01-2222-41, 01-2222-42) stable after staining?

After staining, UltraComp eBeads Compensation Beads (Cat. No. 01-2222-41, 01-2222-42) are stable for up to 3 days if the samples are stored in a fixative. If the samples are stored in FACS buffer, they are stable for 1 week.

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.

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.

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.

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.

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).

Can I use the OneComp and UltraComp eBeads microspheres with Super Bright-conjugated antibodies?

UltraComp eBeads microspheres (Cat. No. 01-2222) are recommended for use with Super Bright dyes.
Note: Super Bright Staining Buffer (Cat. No. SB-4400) is not compatible with UltraComp eBeads microspheres (Cat. No. 01-2222-41, 00-2222-42). If using UltraComp eBeads microspheres as a compensation tool, solely use Flow Cytometry Stain Buffer (Cat. No. 00-4222-26, 00-4222-57) for any antibody dilutions.

In some experiments, we have observed that compensation values for Super Bright 780- and Brilliant Violet 785- or Brilliant Violet 786-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads, both with Super Bright 780 and Brilliant Violet 786. We have not tested Brilliant Violet 785 with the AbC beads.

For Research Use Only. Not for use in diagnostic procedures.