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What are the Super Bright dyes? Product FAQ

Answer

eBioscience Super Bright dyes are a series of patent-pending fluorochromes that are based upon a fluorescent polymer and its tandems. Super Bright dyes have been developed for use in flow cytometry and can be excited by the violet laser (405 nm). The peak emission of each Super Bright dye is indicated by the number in the product description, e.g., "Super Bright 600" has a peak emission of 600 nm.

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Answer Id: E14676

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Can I prepare an antibody cocktail containing Super Bright Staining Buffer and Super Bright-conjugated antibodies ahead of time? Product FAQ

Answer

We recommend that the antibody cocktails containing Super Bright-conjugated antibodies and Super Bright Staining Buffer are prepared fresh prior to staining. Discard any unused portions. We do not recommend overnight storage of prepared cocktails.

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Answer Id: E14688

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Is there a specific buffer I should use when staining with Super Bright-conjugated antibodies? Product FAQ

Answer

No special buffer is required when using a single Super Bright-conjugated antibody in a panel.
When using more than one Super Bright dye, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

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Answer Id: E14680

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Can Super Bright-conjugated antibodies be used in combination with other polymer dyes? What buffer should I use when using more than one Super Bright or polymer dye? Product FAQ

Answer

When using two or more Super Bright dyes, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

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

Answer Id: E14682

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Are the Super Bright dyes sensitive to methanol fixation? Product FAQ

Answer

Super Bright dyes are stable in methanol-based fixation buffers.

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Answer Id: E14686

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Can the Super Bright dyes be used for non-flow applications? Product FAQ

Answer

Super Bright dyes have not been tested for applications other than flow cytometry.

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Answer Id: E14678

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How does the protocol for staining with Super Bright-conjugated antibodies differ from staining with conventional organic fluorochrome and eVolve conjugated antibodies? Product FAQ

Answer

Antibodies conjugated to Super Bright dyes can be used similarly to traditional fluorochromes. If multiple Super Bright dyes are used in combination with each other or in combination with other polymer dyes (i.e., Brilliant Violet dyes), then the use of Super Bright Staining Buffer (Cat. No. SB-4400) is recommended to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

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

Answer Id: E14679

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Are there recommended tubes for staining with Super Bright-conjugated antibodies? Product FAQ

Answer

We recommend using polystyrene tubes (for example, FACS tubes) for staining with your Super Bright-conjugated antibodies. If using polypropylene (for example, Eppendorf tubes), protecting from light is critical.

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Answer Id: E14681

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Which laser do I use to detect Super Bright-conjugated antibodies? Product FAQ

Answer

The violet laser (405 nm) should be used to excite Super Bright-conjugated antibodies.

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Answer Id: E14684

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Can the Super Bright Staining Buffer be used with other polymer dyes? Product FAQ

Answer

Yes, the Super Bright Staining Buffer (Cat. No. SB-4400) is compatible with other polymer dyes (i.e., Brilliant Violet dyes) and is useful for minimizing any non-specific polymer interactions when two or more of these dyes are used in combination.

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

Answer Id: E14683

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What is the difference between the Super Bright and eVolve dyes? Product FAQ

Answer

Super Bright dyes are fluorochromes based on polymer technology and are excited by the violet laser. In contrast, eVolve dyes are based on Qdot nanocrystal technology and are maximally excited by the UV laser, although they may also be excited by violet, blue, yellow-green, and, depending on the eVolve, the red laser lines. The general shape of the emission spectra for Super Bright dyes will be more similar to traditional fluorochromes, while eVolve dyes have extremely narrow emission spectra. Please contact Tech Support (techsupport@thermofisher.com) for more information.

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Answer Id: E14677

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

Answer

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.

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Answer Id: E14689

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Can Super Bright-conjugated antibodies be used in combination with intracellular (IC Fixation/Permeabilization) or intranuclear (Foxp3 fixation/permeabilization) staining? Product FAQ

Answer

Yes, Super Bright-conjugated antibodies are stable in formaldehyde-based fixation buffers and permeabilization buffers, such as the IC Fixation and Permeabilization Buffer Set (Cat. No. 88-8824) and the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523).

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Answer Id: E14685

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Can I fix my cells after staining with Super Bright-conjugated antibodies? How long can fixed cells be stored prior to analysis? Product FAQ

Answer

Samples that have been stained with antibodies conjugated to Super Bright dyes may be stored for up to three days, at 2-8°C, in the dark, using either IC Fixation Buffer (Cat. No. 00-8222) or 1-step Fix/Lyse Buffer (Cat. No. 00-5333) with no significant effect on brightness or compensation.

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Answer Id: E14687

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Are the Super Bright Dyes photo-labile? Product FAQ

Answer

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

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Answer Id: E14441

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