IFN-γ Antibody, anti-human

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

IFN-γ Antibody, anti-humanExtended Validation

Clone:

45-15

Type of antibody:

Primary antibodies, Functional-grade antibodies

Isotype:

mouse IgG1κ, mouse IgG1

Applications:

ICFC, FA

Alternative names:

IFNg, IFG, IFI

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

APC
APC-Vio 770
FITC
PE
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-114-016, 130-113-490

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left images) or stimulated with SEB for six hours. After two hours, brefeldin A was added for stimulated and unstimulated cells. The cells were fixed, permeabilized, and intracellularly stained with Anti-IFN-γ antibodies as well as with CD4 antibodies and CD69 antibodies. Cells were then analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. CD4⁺ lymphocytes were pre-gated for the analysis. Cell debris were excluded from the analysis based on scatter signals.

Functional assay applications forIFN-γ Antibody, anti-human

pure-functional grade

Conjugate:

pure-functional grade

Tested:

during development

Products used:

130-095-743

Type of functional assay:

neutralization of cytokine activity, in vitro inhibition of Tʜ1 cell polarization, promotion of Tʜ2 cell and Tʜ17 cell polarization

Extended validation for IFN-γ Antibody, anti-human

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

Overview

Interferon gamma (IFN-γ) is produced by cells that are involved in inflammatory immune responses. IFN-γ is predominantly released by memory and effector CD4

⁺

and CD8

⁺

T cells as well as by NK cells upon activation. Quantitative analysis of IFN-γ–producing cells can provide important information on the course of immune responses. Anti-IFN‑γ antibodies are designed for intracellular staining of IFN‑γ–producing cells. Cells can be stimulated for IFN-γ–production, e.g. by polyclonal stimulation with mitogens. For induction of IFN‑γ–production by antigen-specific T cells, cells are restimulated with respective antigen. IFN‑γ can be accumulated in the cells by addition of secretion inhibitors like brefeldin A. After fixation and permeabilization of the cell sample, IFN‑γ–producing cells can be stained intracellularly with Anti-IFN‑γ antibodies. Staining of surface markers allows simultaneous flow cytometric analysis of subsets and activation status of the IFN-γ–producing cells.

Alternative names

IFNg, IFG, IFI

Detailed product information

Technical specifications

Clone	45-15
Clonality	monoclonal
Isotype	mouse IgG1κ, mouse IgG1
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies, Functional-grade antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	IFN-γ
Alternative names of antigen	IFNg, IFG, IFI
Molecular mass of antigen [kDa]	16
Entrez Gene ID	3458
RRID	AB_2733589, AB_2733718, AB_2733719, AB_2751182, AB_2751116, AB_2751183, AB_2751117, AB_2733079, AB_2733080, AB_10830868, AB_2733588

Resources for IFN-γ Antibody, anti-human

Documents and Protocols

App notes/customer reports

Efficient and rapid in vitro generation of fully functional multi-virus-specific CD4⁺ and CD8⁺ T cells.

Certificates

Please follow this

link

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

References for IFN-γ Antibody, anti-human

Publications

Chaput, N. et al. (2013)
Phase I clinical trial combining imatinib mesylate and IL-2: HLA-DR
⁺
NK cell levels correlate with disease outcome.
Oncoimmunology 2(2): e23080

Applications

Extended validation