IFN-γ Antibody, anti-human, REAfinity™

Name: IFN-γ Antibody, anti-human, REAfinity™
Availability: InStock

IFN-γ Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA600

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC

Alternative names:

IFN-gamma, Immune interferon

Intracellular flow cytometry applications forIFN-γ Antibody, anti-human, REAfinity™

APC
APC-Vio 770
FITC
PE
PE-Vio 615
PE-Vio 770
Vio B515
Vio Bright V600
Vio R667
Vio R720
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-114-021, 130-113-495

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs), either left unstimulated (left images) or stimulated with 1 µg/mL ionomycin and 20 ng/mL PMA for 2 hours and for additional 4 hours with1 µg/mL brefeldin A at 37 °C, were fixed, permeabilized, and stained with Anti-IFN-γ antibodies as well as with CD69 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Extended validation for IFN-γ 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 REA600
25723	++
25723.11	++
45-15	-
4S.B3	+
B27	++
Cells were incubated with an excess of purified unconjugated IFN-γ (REA600) 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-y. 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 IFN-y antibodies. As a control, IFN-y 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 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 IFN-γ Antibody, anti-human, REAfinity™

Overview

Clone REA600 recognizes the human interferon gamma (IFN-γ) antigen, a cytokine of type II class interferons. IFN-γ is produced by lymphocytes that are involved in inflammatory immune responses. It functions as a potent activator of macrophages, increases the antiviral and antitumor effects of the type I interferons, and has antiproliferative effects on transformed cells. IFN-γ is predominantly released by memory and effector CD4

⁺

and CD8

⁺

T cells as well as by NK cells upon activation.

Additional information: Clone REA600 displays negligible binding to Fc receptors.

Alternative names

IFN-gamma, Immune interferon

Detailed product information

Technical specifications

Clone	REA600
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
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis ) , chimpanzee ( Pan troglodytes ) , baboon, pigtail monkey ( Macaca nemestrina ) , african green monkey ( Chlorocebus aethiops ) , sooty mangabey ( Cercocebus atys )
Antigen	IFN-γ
Alternative names of antigen	IFN-gamma, Immune interferon
Molecular mass of antigen [kDa]	16
Distribution of antigen	macrophages, T cells, NK cells, T cells, NK cells, macrophages
Entrez Gene ID	3458
RRID	AB_2733587, AB_2733716, AB_2733717, AB_2751184, AB_2751118, AB_2726445, AB_2726168, AB_2751185, AB_2751119, AB_2751796, AB_2751736, AB_2652243, AB_2652244, AB_2905266, AB_2905265, AB_2922017, AB_2921999, AB_2922307, AB_2922300, AB_2922306, AB_2922299, AB_2733586

Resources for IFN-γ Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for IFN-γ Antibody, anti-human, REAfinity™

Publications

Gray, P. W. et al. (1982) Structure of the human immune interferon gene. Nature 298(5877): 859-863
Green, J. A. et al. (1969) Immune specific induction of interferon production in cultures of human blood lymphocytes. Science 164(3886): 1415-1417
Schoenborn, J. R. et al. (2007) Regulation of interferon-gamma during innate and adaptive immune responses. Adv. Immunol. 96: 41-101

Applications

Extended validation