UltraComp eBeads series for simplified compensation
Invitrogen eBeads compensation microspheres contain a mixture of antibody-coated positive and uncoated negative beads, combined in one vial. Dispense the eBeads as a single drop for compensation made easy. UltraComp eBeads microspheres were developed with the same benefits as OneComp eBeads microspheres with the added advantage of being optimized for use on the violet laser and added reactive species including rabbit and human. The UltraComp eBeads were designed for ease of use while obtaining the values needed for compensation.
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Browse the table below to understand the differences between the eBeads.
Table 1: Selection guide for compensation eBeads
|UltraComp eBeads Plus compensation beads||UltraComp eBeads compensation beads||OneComp eBeads compensation beads|
|Reactivity||Human, rabbit, hamster, mouse, and rat antibodies with recognition of the kappa and lambda chains.||Hamster, mouse, and rat antibodies with recognition of the kappa and lambda chains.||Hamster, mouse, and rat antibodies with recognition of the kappa and lambda chains.|
|Laser compatibility||Compatible with most standard lasers, UV to 633 nm. Improved for polymer dye use from violet laser.||Compatible with most standard lasers, UV to 633 nm.||Compatible with most standard lasers, not with UV or violet lasers.|
|Size||25 tests | 100 tests||25 tests | 100 tests||25 tests | 100 tests|
|Cat. No||01-3333-41 | 01-3333-42||01-2222-41 | 01-2222-42||01-1111-41 | 01-1111-42|
UltraComp eBeads Plus microspheres
Traditional compensation beads, when used on the violet laser, typically provide poor compensation values when incorporating the increasing number of violet-emitting fluorochromes. UltraComp eBeads Plus were developed to provide a reliably bright signal, resulting in greater accuracy when calculating spillover values.
Advantages of UltraComp eBeads Plus microspheres:
- Optimized for the violet laser with added compensation resolution for Super Bright Polymer Dyes and Brilliant Violet Dyes (Figure 1)
- Supplied in a single vial—one drop contains both antibody-coated positive and uncoated negative beads
- Suitable for multiple species: human, rabbit, hamster, mouse, and rat (Figure 2)
- Recognizes both kappa and lambda chains
Curious on how to use UltraComp eBeads and OneComp? Explore our protocols
Figure 1. Cells were isolated from human whole blood and resuspended in 1X PBS. Beads were vortexed for at least 30 seconds, and one drop of either UltraComp eBeads or UltraComp eBeads Plus was dispensed into a 5 ml Falcon round-bottom polystyrene tube with 1 x 106 cells. Various antibodies in the table shown below were added at 5 uL per sample or manufacturer’s recommended sample volume. Samples were incubated for 20 minutes, protected from light, then washed with 1X PBS + 1% BSA buffer. Each individually stained sample was run on the Attune NxT V6 flow cytometer. For compensation analysis, each file was imported into the FlowJo software, and matrices were generated for cells alone, UltraComp eBeads, or UltraComp eBeads Plus. Each compensation matrix was applied to the cell samples, and data plots were compared.
Figure 2. Staining of UltraComp eBeads Plus with 14 different Invitrogen eFluor 450 dye-conjugated monoclonal antibodies, including one of each subclass commonly used in flow cytometry. Beads were stained with 0.25 µg of each antibody and analyzed by flow cytometry. Each histogram represents one staining antibody.
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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures.