Two-color flow cytometric analysis of NCAM-1 (CD56) expression by fixed and permeabilized human peripheral blood lymphocytes. Erythrocytes were lysed and leucocytes were fixed and permeabilized in a human whole blood sample using 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049; 10 min, 37 ̊ C) followed by BD Phosflow™ Perm Buffer III (Cat. No. 558050; 30 min, on ice). The leucocytes were then stained with BD Horizon™ PE-CF594 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 564963) and BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427) antibodies. The two-color flow cytometric dot plot showing the correlated expression of NCAM-1 (CD56) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of NCAM-1 (CD56) expression on live-stained human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ PE-CF594 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 564963) and BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427) antibodies. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plot showing the correlated expression of NCAM-1 (CD56) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
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Two-color flow cytometric analysis of NCAM-1 (CD56) expression by fixed and permeabilized human peripheral blood lymphocytes. Erythrocytes were lysed and leucocytes were fixed and permeabilized in a human whole blood sample using 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049; 10 min, 37 ̊ C) followed by BD Phosflow™ Perm Buffer III (Cat. No. 558050; 30 min, on ice). The leucocytes were then stained with BD Horizon™ PE-CF594 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 564963) and BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427) antibodies. The two-color flow cytometric dot plot showing the correlated expression of NCAM-1 (CD56) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of NCAM-1 (CD56) expression on live-stained human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ PE-CF594 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 564963) and BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427) antibodies. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plot showing the correlated expression of NCAM-1 (CD56) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Horizon™
N-CAM-1, Neural cell adhesion molecule 1; NKH1; MSK39; Leu-19
Human (QC Testing)
Mouse IgG1, κ
Human NCAM1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
4684
AB_2869631
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- CF™ is a trademark of Biotium, Inc.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. 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. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lyse/Fix Buffer 5X RUO
Size
250 mL
Cat No.
558049
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
564963 Rev. 1
Antibody Details
R19-760
The R19-760 monoclonal antibody specifically binds to NCAM-1 (Neural cell adhesion molecule 1) that is also known as CD56 and NKH1. NCAM-1 is a heavily glycosylated transmembrane protein and a member of the Ig supergene family. NCAM-1 represents an isoform of the neural cell adhesion molecule (NCAM). In the hematopoietic system, it is present on approximately 10% to 25% of peripheral blood lymphocytes. It is expressed on natural killer (NK) cells and a subset of T cells, NKT cells. It is not expressed on myeloid cells, erythrocytes or B cells. This molecule appears to function as an adhesion molecule and can serve as a panspecific NK-cell marker. The R19-760 antibody recognizes an epitope in the NCAM-1 (CD56) extracellular domain that resists destruction due to cellular fixation with BD Phosflow™ Lyse/Fix Buffer and permeabilization with BD Phosflow™ Perm Buffer III.
This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
564963 Rev. 1
Format Details
PE-CF594
BD Horizon™ PE-CF594 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 with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is 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 615 nm (e.g., a 610/20-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 green (532-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.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
564963 Rev.1
Citations & References
View product citations for antibody "564963" on CiteAb
Development References (5)
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Campbell JJ, Qin S, Unutmaz D, et al. Unique subpopulations of CD56+ NK and NK-T peripheral blood lymphocytes identified by chemokine receptor expression repertoire. J Immunol. 2001; 166(11):6477-6482. (Biology). View Reference
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Elsaid AF, Shaheen M, Ghoneum M. Biobran/MGN-3, an arabinoxylan rice bran, enhances NK cell activity in geriatric subjects: A randomized, double-blind, placebo-controlled clinical trial. Exp Ther Med. 2018; 15(3):2313-2320. (Clone-specific: Flow cytometry). View Reference
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Fornara C, Furione M, Arossa A, Gerna G, Lilleri D. Comparative magnitude and kinetics of human cytomegalovirus-specific CD4⁺ and CD8⁺ T-cell responses in pregnant women with primary versus remote infection and in transmitting versus non-transmitting mothers: Its utility for dating primary infection in pregnancy.. J Med Virol. 2016; 88(7):1238-46. (Clone-specific: Flow cytometry). View Reference
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Hsieh CY, Chen CL, Lin YS, et al. Macrophage migration inhibitory factor triggers chemotaxis of CD74+CXCR2+ NKT cells in chemically induced IFN-γ-mediated skin inflammation.. J Immunol. 2014; 193(7):3693-703. (Clone-specific: Flow cytometry). View Reference
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Ritz J, Trinchieri G, Lanier LL. NK-cell Antigens: Section Report. In: 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; 1995:1367-1372.
564963 Rev. 1
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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.