Clone:
REA192
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
ICOS, AILIM, CVID1

Extended validation for CD278 (ICOS) Antibody, anti-human/mouse/rat, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA192
DX29++
ISA-3-
C398.4A++
Cells were incubated with an excess of purified unconjugated CD278 (ICOS) (REA192) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD278 (ICOS). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD278 (ICOS) antibodies and with a suitable counterstaining. As a control, CD278 (ICOS) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD278 (ICOS) (REA192). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD278 (ICOS) (REA192). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD278 (ICOS) (REA192). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD278 (ICOS) Antibody, anti-human/mouse/rat, REAfinity™

Overview

Clone REA192 recognizes CD278 (ICOS), which belongs to the CD28 family of proteins. CD28 family members are costimulatory receptor molecules, contain single immunoglobulin V-like domain, and belong to the immunoglobulin superfamily. CD287 recognizes inducible T cell costimulator ligand (ICOSL) as ligand and its expression is induced on activated T cells. Enhanced expression of CD278 is also observed on T cells, followed by costimulation of T cells via CD28. Further expression of CD278 is found on B cells, monocytes, and dendritic cells. Crosslinking of ICOS on T cell surface in the presence of primary stimulation signal leads to T cell proliferation and differentiation. In addition, CD278 plays role in Tʜ cell differentiation, effector T cell function, and isotype class switching.
Additional information: Clone REA192 displays negligible binding to Fc receptors.

Alternative names

ICOS, AILIM, CVID1

Detailed product information

Technical specifications

CloneREA192
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse, rat
AntigenCD278 (ICOS)
Alternative names of antigenICOS, AILIM, CVID1
Distribution of antigenT cells
RRIDAB_2784101, AB_2801885, AB_2656911, AB_2656912, AB_2656915, AB_2656916, AB_2656917, AB_2656918, AB_2656919, AB_2656920, AB_2656921, AB_2656922, AB_2656923, AB_2656924, AB_2656925, AB_2656926, AB_2656927, AB_2656928, AB_2656929, AB_2656930, AB_2784102

Resources for CD278 (ICOS) Antibody, anti-human/mouse/rat, REAfinity™

References for CD278 (ICOS) Antibody, anti-human/mouse/rat, REAfinity™

Publications

  1. Akbari, O. et al. (2008)
    ICOS/ICOSL interaction is required for CD4
    +
    invariant NKT cell function and homeostatic survival
    J. Immunol. 180: 5448-5456
  2. Wassink, L. et al. (2004) ICOS expression by activated human Tʜ Cells is enhanced by IL-12 and IL-23: Increased ICOS expression enhances the effector function of both Tʜ1 and Tʜ2 cells. J. Immunol. 173: 1779-1786

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