IOTest CD86 (B7-2)-PC7 PN B30648 – 0.5 mL – Liquid – Clone HA5.2B7 Analyte Specific Reagent. Analytical and performance characteristics are not established. SPECIFICITY REAGENT CONTENTS SELECTED RESEARCH REFERENCES The anti-human HA5.2B7 monoclonal This antibody is provided in phosphate- 1. Rennert, P., Furlong, K., Jellis, C., antibody (mAb) binds specifically to CD86 buffered saline, containing 0.1% sodium Greenfield, E., Freeman, G.J., Ueda, Y, (B7-2) antigen (1). azide and 2 mg/mL bovine serum albumin. Levine, B., June, C.H. and Gray, G.S. The CD86 antigen (B7-2, B70) is a single Concentration: See lot specific Certificate of “The IgV domain of human B7-2 chain transmembrane glycoprotein, Analysis at www.beckmancoulter.com. (CD86) is sufficient to co-stimulate T structurally similar to CD80 (B7-1) (2, 3). Its lymphocytes and induce cytokine molecular weight is 80 kDa, under reducing STATEMENTS OF WARNING secretion”, International Immunology, conditions. The extracellular region is 1. This reagent contains 0.1% sodium 1997, 9, 6, 805–813. composed of one V-type and one C-type Ig- azide. Sodium azide under acid 2. Boussiotis, V.A., Freeman, G.J., like domains. There are 8 potential sites for conditions yields hydrazoic acid, an Gribben, J.G., Daley, J., Gray, G., N-glycosylation. The cytoplasmic tail has 3 extremely toxic compound. Azide Nadler, L.M., "Activated human B potential sites for protein kinase C compounds should be flushed with lymphocytes express three CTLA-4 phosphorylation (4, 5). CD86 shares with running water while being discarded. counterreceptors that costimulate T-cell CD80 the same co-receptors on T cells, These precautions are recommended activation", 1993, Proc. Natl. Acad. Sci. CD28 and CD152 (CTLA-4). CD86 binds to to avoid deposits in metal piping in USA, 90, 11059-11063. CD152 with a 20 to 100-fold higher affinity which explosive conditions can 3. Engel, P., Gribben, J.G., Freeman, than to CD28 (6). CD86 and CD80 have a develop. If skin or eye contact occurs, G.J., Zhou, L.J., Nozawa, Y., Abe, M., critical role in one costimulatory pathway wash excessively with water. Nadler, L.M., Wakasa, H., Tedder, T.F., involved in the prevention of antigen-specific 2. Specimens, samples and all material "The B7-2 (B70) costimulatory molecule T-cell tolerance (anergy), mediated by coming in contact with them should be expressed by monocytes and activated ligation of CD28 on T cells by its ligands, considered potentially infectious and B lymphocytes is the CD86 CD86 and CD80 on antigen-presenting cells disposed of with proper precautions. differentiation antigen", 1994, Blood, 5, (7, 8). 3. Never pipet by mouth and avoid 84, 1402-1407. CD86 is constitutively expressed by contact of samples with skin and 4. Hardie, D., L., Casamayor, M., interdigitating dendritic cells, peripheral mucous membranes. Johnson, G.D., Hill, D., Van Schijndel, blood monocytes, activated B cells, and, at a 4. Do not use antibody beyond the G., Van Lier, R., Khan, M., MacLennan, lower level, by peripheral blood dendritic expiration date on the label. I. C.M., "CD86 workshop panel report", cells (2, 3, 9). On lymphocytes, CD86 5. Do not expose reagents to strong light 1997, Leucocyte Typing VI, White Cell appears as a B-cell activation antigen. It is during storage or incubation. Differentiation Antigens. Kishimoto, T., preferentially expressed by memory B cells 6. Avoid microbial contamination of et al, Eds., Garland Publishing, Inc., and germinal B center cells, but not on reagents or incorrect results might 201-204. plasma cells (4). Its expression can be occur. 5. Salamone, M.C., Salamone, G.V., upregulated by activation through surface 7. Use good laboratory practices when Reyes, G.B., Barboza, M., Kado, M., immuno-globulin, MHC-class II molecules, handling this reagent. Ramhorst, R., Fainboim, L., "CD86 and CD40 triggering, or by PMA treatment 8. Any change in the physical appearance workshop: preferential staining of with ionomycin. Also, PHA-activated T cells of the reagents may indicate activated CD8-positive T lymphocytes show a weak expression of CD86 on CD4- deterioration and the reagent should by monoclonal antibody CD86.6", 1997, positive cells, and a consistent one on CD8- not be used. Leucocyte Typing VI, White Cell positive cells (5). STORAGE AND HANDLING CONDITIONS Differentiation Antigens. Kishimoto, T., et al, Eds., Garland Publishing, Inc., REAGENT AND STABILITY This reagent is stable up to the expiration 204-205. IOTest CD86 (B7-2)-PC7 date when stored at 2 – 8°C. Do not freeze. 6. Ellis, J.H., Burden, M.N., Vinogradov, Conjugated Antibody No reconstitution is necessary. This D.V., Linge, C., Crowe, J.S., PN B30648 - 0.5 mL - Liquid monoclonal antibody may be used directly "Interactions of CD80 and CD86 with from the vial. Bring reagent to 18 – 25°C CD28 and CTLA4", 1996, J. Immunol., Clone HA5.2B7 prior to use. 56, 2700-2709. Isotype IgG2b, Mouse 7. Gause, W.C., Halvorson, M.J., Lu, P., Immunogen B7.2 transfected CHO cells PRECAUTIONS Greenwald, R., Linsley, P., Urban, J.F., Hybridoma SP2/0 x balb/c Due to the tandem structure of the Finkelman, F.D., "The function of Source Ascites fluid or supernatant fluorochrome, PC7 also emits light at costimulatory molecules and the of in vitro cultured hybridoma 575 nm. This secondary emission peak development of IL-4 producing T cells", cells. varies from lot-to-lot of PC7. Therefore, for 1997, Immunol. Today, 3, 18, 115-120. Purification Affinity chromatography multi-color analysis, the compensation 8. Dorfman, D., Schultze, J., Shahsafaei, Conjugation R Phycoerythrin-Cyanine 7 matrix should be carefully checked when A., Michalak, S., Gribben, J., Freeman, (PC7) changing the lot of a PC7-conjugate. G., Pinkus, G. and Nadler, L.M, “In Vivo Molar Ratio PC7 / Ig : 0.5 - 1.5 Expression of B7-1 and B7-2 By Fluorescence Excites at 488 nm Follicular Lymphoma Cells Can Prevent Emits at 770 nm Induction of T-Cell Anergy But Is Insufficient to Induce Significant T-Cell Proliferation”, Blood, 1997, 90, 11, 4297-4306. Page 1 of 2 B30648 2013-07-08 IG-720–ASR_L_AN IOTest CD86 (B7-2)-PC7 PN B30648 – 0.5 mL – Liquid – Clone HA5.2B7 9. Ferlazzo, G., Semino, C., Spaggiari, G.M., Meta, M., Mingari, M.C. and , MANUFACTURED BY : Printed in France. Melioli, G. “Dendritic cells efficiently IMMUNOTECH SAS Made in France. cross-prime HLA class I-restricted a Beckman Coulter Company cytolytic T lymphocytes when pulsed 130, avenue de Lattre de Tassigny ©2013 Beckman Coulter, Inc. with both apoptotic and necrotic cells B.P. 177 - 13276 Marseille Cedex 9 but not with soluable cell-derived France lysates”, Int Immunol. 2000, 12, 1741- 1747. For additional information, or if damaged product is received, call Beckman Coulter TRADEMARKS Customer Service at 800-526-7694 (USA or Beckman Coulter, the stylized logo, and Canada) or contact your local Beckman IOTest are trademarks of Beckman Coulter, Coulter Representative. Inc., and registered with the USPTO. www.beckmancoulter.com Page 2 of 2 B30648 2013-07-08 IG-720–ASR_L_AN .