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Flow cytometric analysis of ICOS (CD278) expression on stimulated human peripheral blood lymphocytes. Phytohemagglutinin (PHA)-stimulated (3 days) peripheral blood mononuclear cells were stained with either APC-H7 Mouse Anti-Human ICOS (CD278) antibody (Cat. No. 567142/567143; solid line histogram) or APC-H7 Mouse IgG1 κ Isotype Control (Cat. No. 561427; dashed line histogram). The fluorescence histogram showing ICOS (CD278) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ APC-H7 Mouse Anti-Human ICOS (CD278)
Regulatory Status Legend
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Preparation And Storage
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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
Product Notices
- 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.
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- BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It has decreased intensity but it is engineered for greater stability and less spillover in the APC channel and consequently offers better performance than APC-Cy7. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm. BD recommends that a 750-nm longpass filter be used along with a red-sensitive detector such as the Hamamatsu R3896 PMT. As with APC-Cy7 special filters are required when using APC-H7 in conjunction with APC. Note: Although our APC-H7 products demonstrate higher lot-to lot consistency than other APC tandem conjugate products, and every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-H7 conjugate.
- Although BD APC-H7 is engineered to minimize spillover to the APC channel and is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes, it is still good practice to minimize as much as possible, any light, temperature and fixative exposure when working with all fluorescent conjugates.
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Companion Products
The DX29 monoclonal antibody specifically binds to human CD278, which is also known as Inducible Costimulator (ICOS) or Inducible T-cell Costimulator. ICOS is a homodimeric type I transmembrane glycoprotein with an approximate molecular weight of 50-60 kDa. It is a member of the CD28 family and is highly expressed on activated T cells. CD278 is the receptor for ICOS-ligand (also known as, CD275, B7-H2, B7RP-1, or LICOS). Like CD28, ICOS can provide a costimulatory signal for T cell activation, proliferation and cytokine production. It is not expressed on resting or activated B cells, monocytes, NK cells, granulocytes, dendritic cells or platelets. Unlike the constitutively expressed CD28, ICOS is de novo expressed upon cellular activation. Reports describe similarities between CD28 and ICOS in T cell activation, such as the costimulation of cytokine production. However, it has been suggested that ICOS may play a greater role in IL-10 production. In the presence of IL-10, purified recombinant human ICOS protein significantly increased in vitro B cell growth stimulated by pokeweed mitogen (PWM) and enhanced production of IgG.
Development References (7)
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Ahlmanner F, Sundstrom P, Akeus P, et al. CD39(+) regulatory T cells accumulate in colon adenocarcinomas and display markers of increased suppressive function. Oncotarget. 2018; 9(97):36993-37007. (Clone-specific: Flow cytometry). View Reference
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Aicher A, Hayden-Ledbetter M, Brady WA, et al. Characterization of human inducible costimulator ligand expression and function. J Immunol. 2000; 164(9):4689-4696. (Biology). View Reference
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Dong C, Nurieva RI. Regulation of immune and autoimmune responses by ICOS. J Autoimmun. 2003; 21(3):255-260. (Biology). View Reference
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Fos C, Salles A, Lang V, et al. ICOS ligation recruits the p50alpha PI3K regulatory subunit to the immunological synapse. J Immunol. 2008; 181(3):1969-1977. (Clone-specific: Flow cytometry). View Reference
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Kallinich T, Beier KC, Gelfand EW, Kroczek RA, Hamelmann E. Co-stimulatory molecules as potential targets for therapeutic intervention in allergic airway disease. Clin Exp Allergy. 2005; 35(12):1521-1534. (Biology). View Reference
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Sayin I, Radtke AJ, Vella LA, et al. Spatial distribution and function of T follicular regulatory cells in human lymph nodes. J Exp Med. 2018; 215(6):1531-1542. (Clone-specific: Flow cytometry). View Reference
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Witsch EJ, Peiser M, Hutloff A, et al. ICOS and CD28 reversely regulate IL-10 on re-activation of human effector T cells with mature dendritic cells. Eur J Immunol. 2002; 32(9):2680-2686. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.