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PE Mouse Anti-Human CD59
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PE Mouse Anti-Human CD59
Flow cytometric analysis of CD59 expression on peripheral blood leucocytes. Human or rhesus whole blood was lysed with BD FACS™ Lysing Solution (Cat. No. 349202), then preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 555574; dashed line histogram) or PE Mouse Anti-Human CD59 (Cat. No. 555764/557141/560953; solid line histogram). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of human lymphocytes (Left Panel) or rhesus granulocytes (Right Panel). Flow cytometry was performed on a BD LSRFortessa™ X-20 system.
Flow cytometric analysis of CD59 expression on peripheral blood leucocytes. Human or rhesus whole blood was lysed with BD FACS™ Lysing Solution (Cat. No. 349202), then preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 555574; dashed line histogram) or PE Mouse Anti-Human CD59 (Cat. No. 555764/557141/560953; solid line histogram). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of human lymphocytes (Left Panel) or rhesus granulocytes (Right Panel). Flow cytometry was performed on a BD LSRFortessa™ X-20 system.
Product Details
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BD Pharmingen™
HRF-20; MAC-inhibitory protein; MAC-IP; MACIF; MEM43; MIRL; Protectin; 1F5
Rhesus, Cynomolgus, Baboon (QC Testing), Human (Tested in Development)
Mouse IgG2a, κ
Human Erythrocytes
Flow cytometry (Routinely Tested)
20 µl
V S006
966
AB_2076248
Aqueous buffered solution containing BSA, protein stabilizer, 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 with R-PE under optimum conditions, and unconjugated antibody and free PE were 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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557141 Rev. 7
Antibody Details
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p282 (H19)

The p282 (HI9) monoclonal antibody specifically binds to CD59, a 19 kDa glycosylphosphatidylinositol (GPI)-anchored glycoprotein, expressed on hematopoietic and non-hematopoietic cells. Because of its interaction with complement activated products, CD59 has been termed membrane-attack-complex-inhibitory factor (MACIF), homologus restriction factor (HRF20), membrane inhibitor of reactive lysis (MIRL) and Protectin. It inhibits the cytolytic activity of the complement system by binding to C8 and C9, thereby blocking the assembly of the membrane attack complex. CD59 also participates in spontaneous T-cell/erythrocyte adhesion, interacts with CD2, and plays a role in T-cell activation.

Clone p282 also cross-reacts with peripheral blood leukocytes of baboon and both rhesus and cynomolgus macaque monkeys. The distribution of leukocytes is similar to that observed with peripheral blood leukocytes from normal human donors, with all populations, lymphocytes, monocytes and granulocytes showing reactivity to p282.

557141 Rev. 7
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
557141 Rev.7
Citations & References
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Development References (8)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Davies A, Lachmann PJ. Membrane defence against complement lysis: the structure and biological properties of CD59. Immunol Res. 1993; 12(3):258-275. (Biology). View Reference
  3. Deckert M, Kubar J, Zoccola D, et al. CD59 molecule: a second ligand for CD2 in T cell adhesion. Eur J Immunol. 1992; 22(11):2943-2947. (Biology). View Reference
  4. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  5. Kunii H, Shichishima T, Saitoh Y, Noji H, Maruyama Y. Surface expression of complement regulatory proteins, decay-accelerating factor (DAF) and CD59, on cultured human endothelial cells. Fukushima J Med Sci. 1998; 44(2):69-81. (Biology). View Reference
  6. 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.
  7. Sloand EM, Maciejewski JP, Dunn D, et al. Correction of the PNH defect by GPI-anchored protein transfer. Blood. 1998; 92(11):4439-4445. (Biology). View Reference
  8. Whitlow MB, Iida K, Stefanova I, Bernard A, Nussenzweig V. H19, a surface membrane molecule involved in T-cell activation, inhibits channel formation by human complement. Cell Immunol. 1990; 126(1):176-184. (Biology). View Reference
View All (8) View Less
557141 Rev. 7

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