PE-Cy™5 Rat IgG2b, κ Isotype Control

BD Pharmingen™ PE-Cy™5 Rat IgG2b, κ Isotype Control

Name: PE-Cy™5 Rat IgG2b, κ Isotype Control
Brand: BD Pharmingen™
SKU: 553990

Clone A95-1

(RUO)

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Product Details

Brand: BD Pharmingen™

Reactivity: Mouse (QC Testing)

Isotype: Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2b, κ

Immunogen: TNP-Keyhole Limpet Hemocyanin

Application: Flow cytometry, Isotype control (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_479621

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

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

PE-Cy5™ tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse Fc Block™ (Cat. No. 553141/553142) can reduce the non-specific binding of Cy-Chrome™-conjugated reagents to mouse B cells.

Furthermore, we have observed a distinct type of interaction between PE-Cy5™ tandem fluorochromes and the splenocytes of SJL, NOD, and MRL mice: B lymphocytes and some other leukocyte subsets are brightly stained, and Mouse Fc Block™ has no significant effect. Therefore, Cy-Chrome™-conjugated reagents should not be used to stain leukocytes of SJL, NOD, or MRL mice. Reagents conjugated to PE, PerCP (an alternate "third color" fluorochrome for the Argon ion laser), Allophycocyanin (APC), and PharRed™ (APC-Cy7 tandem fluorochrome) can be used on leukocytes from these mouse strains.

PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5™, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
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.
PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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.
An isotype control should be used at the same concentration as the antibody of interest.

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Antibody Details

A95-1

The A95-1 antibody has unknown specificity. Trinitrophenal (TNP), the immunogen, is a hapten that is not expressed on human, mouse, rat, or non-human primate cells. The A95-1 immunoglobulin was selected as an isotype control following screening for low background on a variety of mouse and human tissues.

553990 Rev. 10

Format Details

PE-Cy5

PE-Cy5 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496 nm and 566 nm and an acceptor dye, Cy™5, with an emission maximum (Em Max) at 670-nm. PE designed to be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 670-nm (e.g., a 670/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627-640-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: PE-Cy5

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 670 nm

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Citations & References

Development References (3)

Takizawa F, Kinet JP, Adamczewski M. Binding of phycoerythrin and its conjugates to murine low affinity receptors for immunoglobulin G. J Immunol Methods. 1993; 162(2):269-272. (Biology). View Reference
Waggoner AS, Ernst LA, Chen CH, Rechtenwald DJ. PE-CY5. A new fluorescent antibody label for three-color flow cytometry with a single laser. Ann N Y Acad Sci. 1993; 677:185-193. (Biology). View Reference
van Vugt MJ, van den Herik-Oudijk IE, van de Winkle JG. Binding of PE-CY5 conjugates to the human high-affinity receptor for IgG (CD64). Blood. 1996; 88(6):2358-2361. (Biology). View Reference

553990 Rev. 10

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.