Multiparameter Flow Cytometric Analysis of CD59 Expression on Human Leucocytes Left Panel: CD59 expression on different leucocyte populations. Whole blood was stained with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; Top Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD59 antibody (Cat. No. 568270; Bottom Plot Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The pseudocolor density plot showing the correlated expression of CD59 [or Ig isotype control] versus side light-scatter signals (SSC) was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Right Panel: CD59 expression on different lymphocyte subpopulations. Whole blood was similarly stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 (Cat. No. 563546/563548) and PE Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 563238) antibodies and with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Top Plots) or Alexa Fluor™ 647 Mouse Anti-Human CD59 antibody (Bottom Plots). Erythrocytes were similarly lysed. The pseudocolor density plots showing the correlated expression of CD59 [or Ig isotype control] versus CD3 (Left Plots) or versus CD56 (Right Plots) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Cell Analyzer System and FlowJo® software.
Multiparameter Flow Cytometric Analysis of CD59 Expression on Human Leucocytes Left Panel: CD59 expression on different leucocyte populations. Whole blood was stained with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; Top Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD59 antibody (Cat. No. 568270; Bottom Plot Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The pseudocolor density plot showing the correlated expression of CD59 [or Ig isotype control] versus side light-scatter signals (SSC) was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Right Panel: CD59 expression on different lymphocyte subpopulations. Whole blood was similarly stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 (Cat. No. 563546/563548) and PE Mouse Anti-Human NCAM-1 (CD56) (Cat. No. 563238) antibodies and with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Top Plots) or Alexa Fluor™ 647 Mouse Anti-Human CD59 antibody (Bottom Plots). Erythrocytes were similarly lysed. The pseudocolor density plots showing the correlated expression of CD59 [or Ig isotype control] versus CD3 (Left Plots) or versus CD56 (Right Plots) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a LSRFortessa™ X-20 Cell Analyzer System and FlowJo® software.
Product Details
BD Pharmingen™
MEM43; MAC-inhibitory protein; Protectin
Human (QC Testing)
Mouse IgG2a, κ
Human Thymocytes and T Lymphocytes
Flow cytometry (Routinely Tested)
5 µl
IV NL 705; V AS S013; BP BP345; T T-103
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 the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- 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.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
565357
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
BUV395 Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
563546
568270 Rev. 2
Antibody Details
MEM-43
The MEM-43 monoclonal antibody specifically recognizes CD59 which is also known as the MEM43 antigen (MEM43). This 19 kDa glycosylphosphatidylinositol (GPI)-anchored glycoprotein is expressed on all hematopoietic and non-hematopoietic cells. Because of its interaction with activated complement products, CD59 has been termed Membrane Attack Complex Inhibition Factor (MACIF), Homologus Restriction Factor (HRF20), Membrane Inhibitor of Reactive Lysis (MIRL), and Protectin. CD59 inhibits complement-mediated cellular lysis by binding to C8 and C9, thereby blocking the final assembly of the complement Membrane Attack Complex (MAC). CD59 also participates in spontaneous T-cell/erythrocyte adhesion, interacts with CD2, and plays a role in T-cell activation.
568270 Rev. 2
Format Details
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 670-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
568270 Rev.2
Citations & References
View product citations for antibody "568270" on CiteAb
Development References (7)
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Bodian DL, Davis SJ, Morgan BP, Rushmere NK. Mutational analysis of the active site and antibody epitopes of the complement-inhibitory glycoprotein, CD59.. J Exp Med. 1997; 185(3):507-16. (Clone-specific). View Reference
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Forsberg UH, Bazil V, Stefanová I, Schröder J. Gene for human CD59 (likely Ly-6 homologue) is located on the short arm of chromosome 11.. Immunogenetics. 1989; 30(3):188-93. (Biology). View Reference
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Horejsí V, Hilgert I, Kristofová H, Bazil V, Bukovský A, Kulhánková J. Monoclonal antibodies against human leucocyte antigens. I. Antibodies against beta-2-microglobulin, immunoglobulin kappa light chains, HLA-DR-like antigens, T8 antigen, T1 antigen, a monocyte antigen, and a pan-leucocyte antigen.. Folia Biol (Praha). 1986; 32(1):12-25. (Immunogen: Flow cytometry). View Reference
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Meri S, Morgan BP, Davies A, et al. Human protectin (CD59), an 18,000-20,000 MW complement lysis restricting factor, inhibits C5b-8 catalysed insertion of C9 into lipid bilayers.. Immunology. 1990; 71(1):1-9. (Clone-specific). View Reference
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Omidvar N, Wang EC, Brennan P, Longhi MP, Smith RA, Morgan BP. Expression of glycosylphosphatidylinositol-anchored CD59 on target cells enhances human NK cell-mediated cytotoxicity.. J Immunol. 2006; 176(5):2915-23. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Sutherland DR, Illingworth A, Marinov I, et al. ICCS/ESCCA consensus guidelines to detect GPI-deficient cells in paroxysmal nocturnal hemoglobinuria (PNH) and related disorders part 2 - reagent selection and assay optimization for high-sensitivity testing.. Cytometry B Clin Cytom. 2018; 94(1):23-48. (Clone-specific: Flow cytometry). View Reference
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Sutherland DR, Keeney M, Illingworth A. Practical guidelines for the high-sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria clones by flow cytometry.. Cytometry B Clin Cytom. 2012; 82(4):195-208. (Clone-specific: Flow cytometry). View Reference
568270 Rev. 2
Please refer to Support Documents for Quality Certificates
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.