PerCP-Cy™5.5 Streptavidin

Consider BD Horizon RealBlue™ 705 (RB705) Reagents, a bright and clean fluorochrome alternative to BD Horizon™ BB700 and PerCP-Cy5.5 More Info

BD Pharmingen™ PerCP-Cy™5.5 Streptavidin

Name: PerCP-Cy™5.5 Streptavidin
Brand: BD Pharmingen™
SKU: 551419

(RUO)

PerCP-Cy5.5 spectra. The absorption spectrum of Streptavidin-PerCP-Cy5.5 is presented in the left panel. The corresponding emission spectrum, at the excitation wavelength of 488 nm, appears in the right panel.

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Scientific Resources

Product Details

Brand: BD Pharmingen™

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_2868934

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. Streptavidin was conjugated with dye under optimum conditions, and unconjugated Streptavidin and free dye were removed

Recommended Assay Procedures

PerCP has been reported to undergo significant photobleaching, the magnitude of which increases as laser power is increased or beam focus is narrowed. For tandem conjugates incorporating PerCP (e.g., PerCP-Cy5.5), the excitation and emission properties of PerCP and the kinetics of energy exchange between the fluorochromes of the tandem dye may limit their effectiveness on high-speed and/or sorting flow cytometers. Therefore, for third-color flow-cytometric analysis using ≥ 25-mW laser power, we recommend PE-Cy5 (formerly BD Cy-Chrome™)-conjugated Streptavidin (Cat. No. 554062).

It is recommended that a 712/20-nm band-pass filter be used with stream-in-air instruments such as the BD FACStar™ and BD FACSVantage™ flow cytometry systems.

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
Cy is a trademark of GE Healthcare.
Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

PE-Cy™5 Streptavidin RUO

Size 0.1 mg Cat No. 554062

551419 Rev. 6

Antibody Details

PerCP-Cy5.5 is a tandem fluorochrome composed of peridinin chlorophyll protein (PerCP), which is excited by the 488-nm line of an Argon ion laser and serves as the energy donor, coupled to the cyanine dye Cy™5.5, which acts as the energy acceptor and fluoresces at 695 nm.

SAv-PerCP-Cy5.5 is a useful second-step reagent for the indirect immunofluorescent staining of cells in combination with biotinylated primary antibodies for flow cytometric analysis. PerCP-Cy5.5 tandem fluorochrome emission is collected in the Fluorescence-3 (FL3) channel of BD FACScan™ and BD FACSCalibur™ flow cytometry systems.

551419 Rev. 6

Format Details

PerCP-Cy5.5

PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PerCP-Cy5.5

Excitation Source: Blue 488 nm

Excitation Max: 482 nm

Emission Max: 676 nm

551419 Rev.6

Citations & References

Development References (2)

Greimers R, Trebak M, Moutschen M, Jacobs N, Boniver J. Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets. Cytometry. 1996; 23(3):205-217. (Biology). View Reference
Shapiro HM. Practical Flow Cytometry, 3rd Edition. New York: Wiley-Liss, Inc; 1995:280-281.

551419 Rev. 6

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

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