PE-iFluor® 594 Tandem

Top) Spectral pattern was generated using a 4-laser spectral cytometer. Spatially offset lasers (355 nm, 405 nm, 488 nm, and 640 nm) were used to create four distinct emission profiles, then, when combined, yielded the overall spectral signature. Bottom) Flow cytometry analysis of whole blood stained with PE/iFlour® 594 anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora flow cytometer in the PE/iFluor® 594 specific B6-A channel.

Stain index comparison of CD4+ signal using fluorophore-labeled antibody conjugates. Human peripheral blood mononuclear cells (PBMCs) were isolated and stained using AAT Bioquest PE/iFluor® 594 anti-human CD4 conjugates or Biolegend PE/Dazzle™ 594 anti-human CD conjugates. The fluorescence signal was monitored using an Aurora flow cytometer in the PE/iFluor® 594 specific B6-A channel.

The fluorescence intensity of PE-iFluor® 594 anti-human CD4 *SK3* conjugate at different concentrations in the range of 0.03125 to 0.5 µg. Results showed that the fluorescence intensity of the CD conjugates remained nearly consistent.

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	~240000
Solvent	Water

Spectral properties

Absorbance (nm)	566
Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	606

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Refrigerated (2-8 °C); Minimize light exposure
UNSPSC	12171501

Overview SDS Protocol

Molecular weight

~240000

Absorbance (nm)

566

Extinction coefficient (cm ^-1 M ^-1)

1960000

Excitation (nm)

565

Emission (nm)

606

Tandem dyes are a unique class of fluorescent molecules that consist of two different covalently linked fluorophores, a donor (e.g. PE or APC) and a longer-wavelength emitting fluorescence acceptor (e.g. Texas Red, Cy5, Cy7, iFluor® 594 or iFluor® 750). PE-iFluor® 594 is a superior replacement to the commonly used PE-Texas Red tandem with significantly improved FRET efficiency and signal/background ratio. Its primary absorption peak is at 565 nm with emission peak around 610 nm. AAT Bioquest also offers a unique preactivated PE-iFluor® 594 to facilitate the PE- iFluor® 594 tandem conjugation to antibodies and other proteins such as streptavidin and other secondary reagents. Our preactivated PE- iFluor® 594 tandem is ready to conjugate, giving much higher yield than the conventionally tedious SMCC-based conjugation chemistry. In addition, our preactivated PE tandems are conjugated to a protein via its amino group that is abundant in proteins while SMCC chemistry targets the thiol group that has to be regenerated by the reduction of antibodies.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	566
Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	606

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
PE-iFluor® 700 Tandem	565	708	1960000
PE-iFluor® 610 Tandem	565	625	1960000
PE-iFluor® 647 Tandem	565	666	1960000
PE-iFluor® 750 Tandem	565	778	1960000
PE-iFluor® 710 Tandem	565	747	1960000
PE-iFluor® 660 Tandem	565	695	1960000
PE-iFluor™ 597 Tandem	565	612	-
PE-iFluor™ 740 Tandem	565	767	1960000
PE-iFluor™ 720 Tandem	565	750	1960000
PE-iFluor™ 770 Tandem	565	795	1960000
Show More (1)

Images

Figure 1. Top) Spectral pattern was generated using a 4-laser spectral cytometer. Spatially offset lasers (355 nm, 405 nm, 488 nm, and 640 nm) were used to create four distinct emission profiles, then, when combined, yielded the overall spectral signature. Bottom) Flow cytometry analysis of whole blood stained with PE/iFlour® 594 anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora flow cytometer in the PE/iFluor® 594 specific B6-A channel.

Figure 2. Stain index comparison of CD4+ signal using fluorophore-labeled antibody conjugates. Human peripheral blood mononuclear cells (PBMCs) were isolated and stained using AAT Bioquest PE/iFluor® 594 anti-human CD4 conjugates or Biolegend PE/Dazzle™ 594 anti-human CD conjugates. The fluorescence signal was monitored using an Aurora flow cytometer in the PE/iFluor® 594 specific B6-A channel.

Figure 3. The fluorescence intensity of PE-iFluor® 594 anti-human CD4 *SK3* conjugate at different concentrations in the range of 0.03125 to 0.5 µg. Results showed that the fluorescence intensity of the CD conjugates remained nearly consistent.

References

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