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BV421 Armenian Hamster Anti-ICOS (CD278)
BV421 Armenian Hamster Anti-ICOS (CD278)
Analysis of ICOS (CD278) Expression Left Panel - Two-color flow cytometric analysis of ICOS (CD278) expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either no antibody (BD Horizon™ BV421 Autofluorescence Control; Top Plot) or BD Horizon BV421 Hamster Anti-ICOS (CD278) antibody (Cat. No. 565886/565887; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour plots showing the correlated expression of ICOS (CD278) [or BD Horizon™ BV421 Autofluorescence] versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Right Panel - Multicolor flow cytometric analysis of ICOS (CD278) expression on mouse thymocytes. Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat anti-Mouse CD4 (Cat. No. 553051/561091) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) antibodies, and either no antibody (BD Horizon BV421 Autofluorescence Control; dashed line histograms) or BD Horizon BV421 Hamster Anti-ICOS (CD278) antibody (solid line histograms). The fluorescence histograms showing ICOS (CD278) expression (or BD Horizon™ BV421 Autofluorescence) were derived from CD4 and CD8 gated events with the forward and side light-scatter characteristics of viable thymocytes as indicated. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Analysis of ICOS (CD278) Expression Left Panel - Two-color flow cytometric analysis of ICOS (CD278) expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either no antibody (BD Horizon™ BV421 Autofluorescence Control; Top Plot) or BD Horizon BV421 Hamster Anti-ICOS (CD278) antibody (Cat. No. 565886/565887; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour plots showing the correlated expression of ICOS (CD278) [or BD Horizon™ BV421 Autofluorescence] versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Right Panel - Multicolor flow cytometric analysis of ICOS (CD278) expression on mouse thymocytes. Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat anti-Mouse CD4 (Cat. No. 553051/561091) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) antibodies, and either no antibody (BD Horizon BV421 Autofluorescence Control; dashed line histograms) or BD Horizon BV421 Hamster Anti-ICOS (CD278) antibody (solid line histograms). The fluorescence histograms showing ICOS (CD278) expression (or BD Horizon™ BV421 Autofluorescence) were derived from CD4 and CD8 gated events with the forward and side light-scatter characteristics of viable thymocytes as indicated. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
ICOS; AILIM; H4; Inducible T-cell costimulator; CVID1; Ly115
Human (QC Testing), Mouse (Tested in Development), Rat, Rhesus (Reported)
Armenian Hamster IgG
Mouse D10.G4.1 T-cell clone
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869726
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565887 Rev. 1
Antibody Details
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C398.4A

The C398.4A monoclonal antibody specifically binds to Inducible Costimulator (ICOS), which is also known as, CD278, Activation-inducible lymphocyte immunomediatory molecule (AILIM), or H4. ICOS is a type I transmembrane glycoprotein that forms a disulfide-linked homodimer and belongs to the CD28 family within the Ig superfamily. ICOS is expressed on either CD4+ or CD8+ single-positive mature thymocytes, T cells, or subsets of Innate Lymphoid Cells (ILC). Its expression is upregulated on activated T lymphocytes. ICOS is a costimulatory receptor that can bind to the B7-H2 ligand (CD275, ICOS-L) that is expressed on B cells, monocytes, macrophages, dendritic cells, and endothelial cells. ICOS plays a critical role in many types of T cell-dependent immunity. In the case of humoral immunity, for example, ICOS signaling is critical for the differentiation of T follicular helper (Tfh) cells and development of germinal centers. Although C398.4A was generated against mouse ICOS, this antibody reportedly crossreacts with human, rhesus, and rat ICOS.

565887 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565887 Rev.1
Citations & References
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View product citations for antibody "565887" on CiteAb

Development References (8)

  1. Araujo LM, Fert I, Jouhault Q, et al. Increased production of interleukin-17 over interleukin-10 by treg cells implicates inducible costimulator molecule in experimental spondyloarthritis. Arthritis Rheum. 2014; 66(9):2412-2422. (Clone-specific: Flow cytometry). View Reference
  2. Brenchley JM, Vinton C, Tabb B, et al. Differential infection patterns of CD4+ T cells and lymphoid tissue viral burden distinguish progressive and nonprogressive lentiviral infections.. Blood. 2012; 120(20):4172-81. (Clone-specific: Flow cytometry). View Reference
  3. Buonfiglio D, Bragardo M, Bonissoni S, et al. Characterization of a novel human surface molecule selectively expressed by mature thymocytes, activated T cells and subsets of T cell lymphomas. Eur J Immunol. 1999; 29(9):2863-2879. (Clone-specific: Flow cytometry, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay). View Reference
  4. Buonfiglio D, Bragardo M, Redoglia V, et al. The T cell activation molecule H4 and the CD28-like molecule ICOS are identical. Eur J Immunol. 2000; 30(12):3463-3467. (Clone-specific: Blocking, Flow cytometry). View Reference
  5. Chen Y, Shen S, Gorentla BK, Gao J, Zhong XP. Murine regulatory T cells contain hyperproliferative and death-prone subsets with differential ICOS expression. J Immunol. 2012; 188(4):1698-1707. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  6. Klatt NR, Vinton CL, Lynch RM et al. SIV infection of rhesus macaques results in dysfunctional T- and B-cell responses to neo and recall Leishmania major vaccination. Blood. 2011; 118(22):5803-5812. (Clone-specific: Flow cytometry). View Reference
  7. Redoglia V, Dianzani U, Rojo JM, et al. Characterization of H4: a mouse T lymphocyte activation molecule functionally associated with the CD3/T cell receptor. Eur J Immunol. 1996; 26(11):2781-2789. (Immunogen: Bioassay, (Co)-stimulation, Flow cytometry, Functional assay, Immunoprecipitation, Radioimmunoassay). View Reference
  8. Xu H, Wang X, Lackner AA, Veazey RS. PD-1(HIGH) Follicular CD4 T Helper Cell Subsets Residing in Lymph Node Germinal Centers Correlate with B Cell Maturation and IgG Production in Rhesus Macaques. Front Biosci. 2014; 5(85):1-7. (Clone-specific: Flow cytometry). View Reference
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565887 Rev. 1

 

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