Clone:
REA1063
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
CTLA-8, IL-17

Extended validation for IL-17A Antibody, anti-human, 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 REA1063
CZ8-23G1++
BL168++
N49-653++
eBio64DEC17-
Cells were incubated with an excess of purified unconjugated IL-17A (REA1063) 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. 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 IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for IL-17A. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with IL-17A antibodies. As a control, IL-17A antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for IL-17A Antibody, anti-human, REAfinity™

Overview

Clone REA1063 recognizes human interleukin 17A (IL-17A). IL-17A, also known as CTLA8, is a member of the IL-17 family (IL-17A–F), is a disulfide-linked homodimeric glycoprotein. IL-17A is produced by CD4
+
T helper (Tʜ) cells, a third T cell subset termed Tʜ17, which secrete also cytokines such as IL-17F and IL-22 and express the natural killer (NK) cell marker CD161. IL-17A secretion has also been described for other cell types, such as CD8
+
memory T cells. Furthermore, intracellular IL-17A has also been detected in eosinophils, neutrophils, and blood monocytes. Emerging data about Tʜ17 cells suggest that these cells are involved in the recruitment of neutrophils to control early stages of infections to a number of pathogens, such as extracellular bacteria and fungi. IL-17A and Tʜ17 cells have been shown to play an important role in immune-mediated diseases, such as rheumatoid arthritis, psoriasis, multiple sclerosis, asthma, inflammatory bowel diseases, and other immune-mediated inflammatory conditions.
Depending on the cytokine milieu present at time of the initial engagement, CD4
+
naive T cells can differentiate into various subsets (Tʜ1, Tʜ2, and Tʜ17). For the differentiation into Tʜ17 cells several cytokines have been described, including TGF-β, IL-1β, IL-6, IL-21, and IL-23. RORγt was identified as the master regulator gene for Tʜ17 cells.
Additional information: Clone REA1063 displays negligible binding to Fc receptors.

Alternative names

CTLA-8, IL-17

Detailed product information

Technical specifications

CloneREA1063
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenIL-17A
Alternative names of antigenCTLA-8, IL-17
Molecular mass of antigen [kDa]15
Distribution of antigenmonocytes, T cells, CD4+ T cells, CD8+ T cells, eosinophils, neutrophils, Th17 cells
Entrez Gene ID3605
RRIDAB_2751465, AB_2751500, AB_2751466, AB_2751501, AB_2751467, AB_2751502, AB_2751468, AB_2751503, AB_2751469, AB_2751504, AB_2751470, AB_2751505, AB_2751471, AB_2751499

References for IL-17A Antibody, anti-human, REAfinity™

Publications

  1. Yao, Z. et al. (1995) Human IL-17: a novel cytokine derived from T cells. J. Immunol. 155: 5483-5486
  2. Kolls, J. K. and Linden, A. (2004) Interleukin-17 family members and inflammation. Immunity 21: 467-476
  3. Ivanov, I. I. et al. (2006)
    The orphan nuclear receptor RORγt directs the differentiation program of proinflammatory IL-17
    +
    T helper cells.
    Cell 126: 1121-1133
  4. Cosmi, L. et al. (2008)
    Human interleukin 17–producing cells originate from a CD161
    +
    CD4
    +
    T cell precursor.
    J. Exp. Med. 205: 1903-1916
  5. Manel, H. et al. (2008) The differentiation of human Tʜ-17 cells requires transforming growth factor-β and induction of the nuclear receptor RORγt. Nat. Immunol. 9: 641-649
  6. Tesmer, L. A. et al. (2008) Tʜ17 cells in human disease. Immunol. Rev. 223: 87-113
  7. Volpe, E. et al. (2008) A critical function for transforming growth factor-β, interleukin 23 and proinflammatory cytokines in driving and modulating human Tʜ-17 responses. Nat. Immunol. 9: 650-657
  8. Yang, L. et al. (2008) IL-21 and TGF-beta are required for differentiation of human Tʜ17 cells. Nature 454: 350-352

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