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Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56)
Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56)
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 Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 563443) and PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of CD3 versus NCAM-1 (CD56) for gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56)
Two-color flow cytometric analysis of NCAM-1 (CD56) expression on live-stained human peripheral blood lymphocytes. Whole blood was stained with Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56) and PE Mouse Anti-Human CD3 antibodies. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plot shows the correlated expression patterns of CD3 versus NCAM-1 (CD56) for gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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 Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 563443) and PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) antibodies. The two-color flow cytometric dot plot shows the correlated expression patterns of CD3 versus NCAM-1 (CD56) for gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of NCAM-1 (CD56) expression on live-stained human peripheral blood lymphocytes. Whole blood was stained with Alexa Fluor® 647 Mouse Anti-Human NCAM-1 (CD56) and PE Mouse Anti-Human CD3 antibodies. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plot shows the correlated expression patterns of CD3 versus NCAM-1 (CD56) for gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
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), Flow cytometry (Tested During Development)
5 µl
4684
AB_2738209
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 monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
563443 Rev. 1
Antibody Details
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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.

563443 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563443 Rev.1
Citations & References
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Development References (14)

  1. Bennett IM, Zatsepina O, Zamai L, Azzoni L, Mikheeva T, Perussia B. Definition of a natural killer NKR-P1A+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12. J Exp Med. 1996; 184(5):1845-1856. (Biology). View Reference
  2. 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
  3. Cooper MA, Fehniger TA, Caligiuri MA. The biology of human natural killer–cell subsets. Trends Immunol. 2001; 22(11):633-640. (Biology). View Reference
  4. Cunningham BA, Hemperly JJ, Murray BA, Prediger EA, Brackenbury R, Edelman GM. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science. 1987; 236(4803):799-806. (Biology). View Reference
  5. Edelman GM. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986; 2:81-116. (Biology). View Reference
  6. Galandrini R, Tassi I, Mattia G, et al. SH2-containing inositol phosphatase (SHIP-1) transiently translocates to raft domains and modulates CD16-mediated cytotoxicity in human NK cells. Blood. 2001; 100(13):4581-4589. (Biology). View Reference
  7. Gerosa F, Baldani-Guerra B, Nisii C, Marchesini V, Carra G, Trinchieri G. Reciprocal activating interaction between natural killer cells and dendritic cells. J Exp Med. 2002; 195(3):327-333. (Biology). View Reference
  8. Lanier LL, Chang C, Azuma M, Ruitenberg JJ, Hemperly JJ, Phillips JH. Molecular and functional analysis of human natural killer cell-associated neural cell adhesion molecule (N-CAM/CD56). J Immunol. 1991; 146(12):4421-4426. (Biology). View Reference
  9. Lanier LL, Le AM, Civin CI, Loken MR, Phillips JH. The relationship of CD16 (Leu-11) and Leu-19 (NKH-1) antigen expression on human peripheral blood NK cells and cytotoxic T lymphocytes. J Immunol. 1986; 136(12):4480-4486. (Biology). View Reference
  10. Lanier LL, Testi R, Bindl J, Phillips JH. Identity of Leu-19 (CD56) leukocyte differentiation antigen and neural cell adhesion molecule. J Exp Med. 1989; 169(6):2233-2238. (Biology). View Reference
  11. Nitta T, Yagita H, Sato K, Okumura K. Involvement of CD56 (NKH-1/Leu-19 antigen) as an adhesion molecule in natural killer–target cell interaction. J Exp Med. 1989; 170(5):1757-1761. (Biology). View Reference
  12. Phillips JH, Lanier LL. Dissection of the lymphokine-activated killer phenomenon: relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986; 164(3):814-825. (Biology). View Reference
  13. 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.
  14. Schubert W, Zimmermann K, Cramer M, Starzinski-Powitz A. Lymphocyte antigen Leu-19 as a molecular marker of regeneration in human skeletal muscle. Proc Natl Acad Sci U S A. 1989; 86(1):307-311. (Biology). View Reference
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563443 Rev. 1

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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.