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PE-Cy™5 Mouse IgG2a, κ Isotype Control
商品详情
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BD Pharmingen™
IgG2a kappa Isotype Control
Mouse BALB/c IgG2a, κ
TNP-keyhole limpet hemocyanin
Flow cytometry, Isotype control (Routinely Tested), Intracellular staining (flow cytometry) (Not Recommended)
0.2 mg/ml
AB_479606
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


准备和存储

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.

推荐的实验流程

Note:  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 BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4 G2 (Cat. No. 553141/553142) can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells.

Furthermore, we have observed a distinct type of interaction between PE-Cy5 tandem fluorochromes and splenocytes of SJL, NOD, and MRL mice: B lymphocytes and some other leukocytes subsets are brightly stained, and Mouse BD Fc Block™ has no significant effect. Therefore, PE-Cy5-conjugated should not be used to stain leukocytes of SJL, NOD, or MRL mice. Reagents conjugated to PE, PerCP, PerCP-Cy5.5, Allophycocyanin (APC), and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.

商品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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.
  5. 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™.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Cy is a trademark of GE Healthcare.
  9. 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 BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
553458 Rev. 18
抗体详情
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G155-178

The G155-178 clone has an unknown specificity. Trinitrophenol (TNP), the immunogen, is a hapten not expressed on human, mouse, rat or non-human primate cells. In the absence of specific binding, this antibody may bind non-specifically to immunoglobulin Fc receptors. The immunoglobulin secreted by the G155-178 hybridoma was selected as a mouse IgG2a, κ isotype control following screening for low background binding on a variety of mouse and human tissues.

553458 Rev. 18
格式详情
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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.
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PE-Cy5
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
670 nm
553458 Rev.18
报价单和参考
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研发参考 (3)

  1. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Biology). 查看参考
  2. 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). 查看参考
  3. 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). 查看参考
553458 Rev. 18

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