IL-17F Antibody, anti-human, REAfinity™

Name: IL-17F Antibody, anti-human, REAfinity™
Availability: InStock

IL-17F Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA1012

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC

Alternative names:

Interleukin-17F, CANDF6, ML-1

Intracellular flow cytometry applications forIL-17F Antibody, anti-human, REAfinity™

APC
FITC
PE
Vio R667

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-116-990, 130-116-870

Protocols:

Intracellular flow cytometry staining protocol (Inside Stain Kit 1:50)

Description:

CD4⁺ T cells were isolated from human peripheral blood mononuclear cells (PBMCs). Cells were then stimulated with 50 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 6 hours and then positively selected for their IL-17A expression. After cultivation for 11 days, cells were restimulated with 50 ng/mL PMA and 1 µg/ml Ionomycin for 6 hours, followed by an incubation with 1 µg/ml Brefeldin A for 4 hours. Cells were then fixed, permeabilized and stained with the REA Control (I) antibodes (left image) or with Anti-IL-17F antibodies. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

Extended validation for IL-17F 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 clones	Overlap in epitope recognition with REA1012
O33-782	++
SHLR17	++
LN2-9C4	++
Cells were incubated with an excess of purified unconjugated IL-17F (REA1012) 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.

Flow cytometric comparison of different clones for IL-17F. 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 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-17F antibodies. As a control, IL-17F antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD3⁺ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye 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 IL-17F. 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 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-17F antibodies. As a control, IL-17F antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD3⁺ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye 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 IL-17F. 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 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-17F antibodies. As a control, IL-17F antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD3⁺ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye 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 IL-17F. 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 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-17F antibodies. As a control, IL-17F antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD3⁺ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye 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 IL-17F. 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 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-17F antibodies. As a control, IL-17F antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD3⁺ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

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

Overview

Clone REA1012 recognizes the human interleukin 17F (IL-17F) antigen, a homodimeric, 34 kDa member of the IL-17 family, which includes IL-17A, IL-17B, IL-17C, IL-17D, and IL-17E (also called IL-25). Interleukin 17 functions as a proinflammatory cytokine that responds to the invasion of the immune system by extracellular pathogens and induces destruction of the pathogen’s cellular matrix. Phylogenetic analysis reveals that among IL-17 family members, the IL-17F isoforms 1 and 2 (ML-1) have the highest homology to IL-17A (sharing 55 and 40% amino acid identity to IL-17A). IL-17F has been shown to have a pro-inflammatory role in asthma. It is expressed in the airway of asthmatics and its expression level is correlated with disease severity. IL-17F is able to induce several cytokines, chemokines and adhesion molecules in bronchial epithelial cells, vein endothelial cells, fibroblasts, and eosinophils. It is derived from several cell types such as T helper 17 (Tʜ17) cells, mast cells and basophils, and shows a wide tissue expression pattern including lung. Overexpression of the IL-17F gene in the airway of mice is associated with airway neutrophilia, the induction of many cytokines, an increase in airway hyperreactivity, and mucus hypersecretion. Hence, IL-17F may have a crucial role in allergic airway inflammation, and have important therapeutic implications in asthma.

Additional information: Clone REA1012 displays negligible binding to Fc receptors.

Alternative names

Interleukin-17F, CANDF6, ML-1

Detailed product information

Technical specifications

Clone	REA1012
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	IL-17F
Alternative names of antigen	Interleukin-17F, CANDF6, ML-1
Molecular mass of antigen [kDa]	15
Distribution of antigen	granulocytes, mast cells, T cells, basophils
Entrez Gene ID	112744
RRID	AB_2727728, AB_2727799, AB_2727729, AB_2727800, AB_2727730, AB_2727801, AB_2727731, AB_2727798

Resources for IL-17F Antibody, anti-human, REAfinity™

Certificates

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

Publications

Starnes, T. et al. (2001) Cutting edge: IL-17F, a novel cytokine selectively expressed in activated T cells and monocytes, regulates angiogenesis and endothelial cell cytokine production. J. Immunol. 167(8): 4137-4140
Yang, X. O. et al. (2008) Regulation of inflammatory responses by IL-17F. J. Exp. Med. 205(5): 1063-1075
Kawaguchi, M. et al. (2009) Role of interleukin-17F in asthma. Inflamm. Allergy Drug Targets 8(5): 383-389

Applications

Extended validation