Anti-Hu CD59 Alexa Fluor® 647

*IMPORTANT ARTICLE: Robert Sutherland D, Keeney M, Illingworth A: Practical guidelines for the high-sensitivity detection and monitoring of paroxysmal nocturnal hemoglobinuria (PNH) clones by flow cytometry. Cytometry B Clin Cytom. 2012 Apr 12. doi: 10.1002/cyto.b.21023. [Epub ahead of print]. This article recommends PE-conjugated MEM-43 as a good reagent for red blood cell analysis of PNH (Paroxysmal Nocturnal Hemoglobinuria) by flow cytometry., URL: https://www.ncbi.nlm.nih.gov/pubmed/22499484, *Horejsi V, Hilgert I, Kristofova H, Bazil V, Bukovsky A, Kulhankova 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. (original description of MEM-43 antigen), URL: https://www.ncbi.nlm.nih.gov/pubmed/2422063?dopt=A, *Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989) ., URL: , *Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995) ., URL: , *Forsberg UH, Bazil V, Stefanova I, Schroder J: Gene for human CD59 (likely Ly-6 homologue) is located on the short arm of chromosome 11. Immunogenetics. 1989;30 (3) :188-93., URL: https://www.ncbi.nlm.nih.gov/pubmed/2476389?dopt=A, *Stefanova I, Hilgert I, Kristofova H, Brown R, Low MG, Horejsi V: Characterization of a broadly expressed human leucocyte surface antigen MEM-43 anchored in membrane through phosphatidylinositol. Mol Immunol. 1989 Feb;26 (2) :153-61., URL: https://www.ncbi.nlm.nih.gov/pubmed/2918859?dopt=A, *Stefanova I, Horejsi V, Ansotegui IJ, Knapp W, Stockinger H: GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254 (5034) :1016-9., URL: https://www.ncbi.nlm.nih.gov/pubmed/1719635?dopt=A, *Cinek T, Horejsi V: The nature of large noncovalent complexes containing glycosyl-phosphatidylinositol-anchored membrane glycoproteins and protein tyrosine kinases. J Immunol. 1992 Oct 1;149 (7) :2262-70., URL: https://www.ncbi.nlm.nih.gov/pubmed/1382093?dopt=A, *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 Feb 3;185 (3) :507-16., URL: https://www.ncbi.nlm.nih.gov/pubmed/9053451?dopt=A, *Cebecauer M, Cerny J, Horejsi V: Incorporation of leucocyte GPI-anchored proteins and protein tyrosine kinases into lipid-rich membrane domains of COS-7 cells. Biochem Biophys Res Commun. 1998 Feb 24;243 (3) :706-10., URL: https://www.ncbi.nlm.nih.gov/pubmed/9500981?dopt=A, *Ilangumaran S, Briol A, Hoessli DC: CD44 selectively associates with active Src family protein tyrosine kinases Lck and Fyn in glycosphingolipid-rich plasma membrane domains of human peripheral blood lymphocytes. Blood. 1998 May 15;91 (10) :3901-8., URL: https://www.ncbi.nlm.nih.gov/pubmed/9573028?itool=, *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 Mar 1;176 (5) :2915-23., URL: https://www.ncbi.nlm.nih.gov/pubmed/16493049?ordin, *Sutherland DR, Richards SJ, Ortiz F, Nayyar R, Benko M, Marinov I, Illingworth A: CD71 improves delineation of PNH type III, PNH type II, and normal immature RBCS in patients with paroxysmal nocturnal hemoglobinuria. Cytometry B Clin Cytom. 2019 Nov 8. doi: 10.1002/cyto.b.21853. , URL: https://www.ncbi.nlm.nih.gov/pubmed/31705743, *Sutherland DR, Illingworth A, Marinov I, Ortiz F, Andreasen J, Payne D, Wallace PK, Keeney M: 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 Jan;94 (1) :23-48., URL: https://www.ncbi.nlm.nih.gov/pubmed/29236353

*Baalasubramanian S, Harris CL, Donev RM, Mizuno M, Omidvar N, Song WC, Morgan BP: CD59a is the primary regulator of membrane attack complex assembly in the mouse. J Immunol. 2004 Sep 15;173 (6) :3684-92., URL: https://www.ncbi.nlm.nih.gov/pubmed/15356114?ordin, *Menu E, Tsai BC, Bothwell AL, Sims PJ, Bierer BE: CD59 costimulation of T cell activation. CD58 dependence and requirement for glycosylation. J Immunol. 1994 Sep 15;153 (6) :2444-56., URL: https://www.ncbi.nlm.nih.gov/pubmed/7521361?ordina, *Rooney IA, Davies A, Griffiths D, Williams JD, Davies M, Meri S, Lachmann PJ, Morgan BP: The complement-inhibiting protein, protectin (CD59 antigen), is present and functionally active on glomerular epithelial cells. Clin Exp Immunol. 1991 Feb;83 (2) :251-6., URL: https://www.ncbi.nlm.nih.gov/pubmed/1704296?ordina, *Meri S, Morgan BP, Davies A, Daniels RH, Olavesen MG, Waldmann H, Lachmann PJ: 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 Sep;71 (1) :1-9., URL: https://www.ncbi.nlm.nih.gov/pubmed/1698710?ordina

The antibody MEM-43 reacts with well defined epitope (W40, R-53) on CD59 (Protectin), an 18-20 kDa glycosylphosphatidylinositol (GPI) -anchored glycoprotein expressed on the surface of all hematopoietic cells; it is widely present on cells in all tissues. This antibody does not compete with MEM-43/5, and has blocking activity.