BB700 Mouse Anti-Human CD123

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

BD Horizon™ BB700 Mouse Anti-Human CD123

Name: BB700 Mouse Anti-Human CD123
Brand: BD Horizon™
SKU: 566483

Clone 7G3

(RUO)

Multiparameter flow cytometric analysis of CD123 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BB700 Mouse IgG2a, κ Isotype Control (Cat. No. 566419; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD123 antibody (Cat. No. 566482/566483; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

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Product Details

Brand: BD Horizon™

Alternative Name: IL3RA; IL-3RA; IL-3Rα; IL-3R-alpha; Interleukin-3 receptor subunit alpha

Reactivity: Human (QC Testing)

Isotype: Mouse IgG2a, κ

Immunogen: Human IL-3Ra-transfected cells

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Entrez Gene ID: 3563

RRID: AB_2869770

Storage Buffer: Aqueous buffered solution containing ≤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. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
An isotype control should be used at the same concentration as the antibody of interest.
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.
The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
Cy is a trademark of GE Healthcare.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Lysing Solution 10X Concentrate CE_IVD

Size 100 mL Cat No. 349202

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Antibody Details

7G3

The 7G3 monoclonal antibody specifically recognizes human CD123, the 70 kDa IL-3 Receptor α (IL-3Rα) chain. CD123 associates with CD131, the 120-140 kDa Common β chain to form the IL-3 Receptor Complex. CD131 is shared with the receptors for interleukins IL-5 and GM-CSF. IL-3Rα is expressed on hematopoietic progenitors and plays an important role in hematopoietic progenitor cell growth and differentiation. It is also expressed by mast cells, macrophages and a CD5+ B cell subset. This antibody has been reported to block the binding of 125I-IL-3 to high and low affinity IL-3 receptors. In functional experiments, this antibody was found to inhibit acute myeloid leukemia cell proliferation, basophil histamine release, endothelial cell-mediated IL-8 secretion, and neutrophil transmigration. This antibody has been reported to be useful for immunoprecipitation, Western blot and immunofluorescent staining for flow cytometry. At the Fifth HLDA Workshop, the human IL-3 receptor was designated CD123.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

Format Details

BB700

The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BB700

Excitation Source: Blue 488 nm

Excitation Max: 476 nm

Emission Max: 695 nm

Citations & References

Development References (5)

Korpelainen EI, Gamble JR, Smith WB, et al. The receptor for interleukin 3 is selectively induced in human endothelial cells by tumor necrosis factor alpha and potentiates interleukin 8 secretion and neutrophil transmigration.. Proc Natl Acad Sci USA. 1993; 90(23):11137-41. (Biology). View Reference
Macardle PJ, Chen Z, Shih CY, et al. Characterization of human leucocytes bearing the IL-3 receptor. Cell Immunol. 1996; 168(1):59-68. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
Smith WB, Guida L, Sun Q, et al. Neutrophils activated by granulocyte-macrophage colony-stimulating factor express receptors for interleukin-3 which mediate class II expression. Blood. 1995; 86(10):3938-3944. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
Sun Q, Woodcock JM, Rapoport A, et al. Monoclonal antibody 7G3 recognizes the N-terminal domain of the human interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3 receptor antagonist.. Blood. 1996; 87(1):83-92. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Neutralization). View Reference

View All (5)

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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.