CD45R (B220) Monoclonal Antibody (RA3-6B2), eVolve™ 655, eBioscience™ - FAQs

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13 个常见问题解答

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

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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Can eVolve-conjugated antibodies be used in combination with intracellular staining?

Yes! eVolve conjugates were designed to be compatible with eBioscience's intracellular staining buffers including the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523-00) and the Intracellular Fixation and Permeabilization Buffer Set (Cat. No. 88-8824-00).

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Where can I purchase specific filter sets suitable for use with eVolve-conjugated antibodies?

Filter sets can be purchased from Chroma Technology, Omega Optical, or Semrock, Inc.

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Which laser do I use to detect eVolve conjugated antibodies?

eVolve-conjugated antibodies are excited by the 355 nm (UV), 405 nm (Violet) and to a lesser extent the 488 nm, 532 nm, and 561 nm lasers. We recommend using the 355 nm or 405 nm laser for optimal performance.

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How does the protocol for staining with eVolve-conjugated antibodies differ from staining with conventional organic fluorochromes or eFluor NC-conjugated antibodies?

The eVolve-conjugated antibodies may be used in the same fashion as other fluorochromes with routine staining in PBS-based buffers. Antibodies conjugated to eVolves are pre-titrated and diluted to the optimal concentration for ease of use. Please refer to the product Technical Data Sheet.

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Which channels will require compensation with eVolve 605- and 655-conjugated antibodies?

In general, the emission spectra of eVolve-conjugated antibodies are narrow, resulting in minimal spectral overlap with other detectors and thus requiring minimal compensation. The filters and instrument settings used in other detectors will determine the amount of compensation that may be necessary. Please be aware that eVolve 655 will require compensation out of the APC channel.

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What is the best application for eVolve fluorophores?

eVolve fluorophores are very bright with narrow emission peaks and, as such, are a beautiful addition to almost any multicolor flow cytometry panel. They are not, however, suitable for IHC applications.

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Are eVolve fluorophores the same as eFluor nanocrystals?

No. While some of the methods to generate eVolve dyes were adapted from nanocrystals, eVolve dyes are unique and free of many of the limitations of previous nanocrystal technologies.

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What are eVolve fluorophores?

eVolve fluorophores were developed as the next logical step after the nanocrystal. eVolves combine the excellent optical properties of QDot nanocrystals with the conjugation expertise of eBioscience to generate a conjugate without the limitations of previous nanocrystal technologies.

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What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

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What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

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How do I dispose of eVolve conjugates?

The eVolve fluorophores are nanoparticles composed of a Cadmium core. We recommend their disposal in accordance with your local, state, and federal regulations for reagents containing heavy metals. Please contact your institution's health and safety officers in order to ascertain the proper disposal of this material. For further information, please contact techsupport@thermofisher.com, to request the Material Safety Data Sheet (MSDS) for these products.

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Are eBioscience eVolve dyes toxic?

All eVolve dyes contain trace amounts of the heavy-metal Cadmium (Cd), in addition to Zinc (Zn) and Selenium (Se). Cadmium is potentially harmful and should be handled with gloves. We recommend that eVolve dyes be handled in accordance with laboratory practices for heavy metals and that users wear appropriate personal protective equipment at all times. These low levels of Cadmium are not likely to pose health risks, however prolonged exposure studies have not been conducted. For further information, please contact techsupport@thermofisher.com, to request the Material Safety Data Sheet (MSDS) for these products.

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