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IFN gamma Monoclonal Antibody (4S.B3), PE-eFluor™ 610, eBioscience™
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FIGURE: 1 / 84
IFN gamma Antibody (61-7319-41) in Flow
Normal human peripheral blood cells were unstimulated (left) or stimulated with Cell Stimulation Cocktail (plus protein transport inhibitors) (500X) (Product # 00-4975-93) (right). Cells were fixed and stained intracellularly with Anti-Human CD4 APC (Product # 17-0047-42) and Anti-Human IFN gamma PE-eFluor® 610, using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis.
Product Details
61-7319-41
Applications
Tested Dilution
Publications
Product Specifications
Published species
Mouse
Host/Isotype
Mouse
/ IgG1, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
4S.B3
Conjugate
Excitation/Emission Max
565/606 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
Product Specific Information
Description: The 4S.B3 monoclonal antibody reacts with interferon-gamma (IFN gamma). Human IFN gamma is a 17 kDa factor produced by activated T and NK cells and is an anti-viral and anti-parasitic cytokine. IFN gamma in synergy with other cytokines, such as TNF alpha, inhibits proliferation of normal and transformed cells. Immunomodulatory effects of IFN gamma are exerted on a wide range of cell types expressing the high affinity receptors for IFN gamma. Glycosylation of IFN gamma does not affect its biological activity.
Applications Reported: This 4S.B3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This 4S.B3 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
IFN gamma (Interferon gamma, Type II interferon) is a macrophage activation factor, and immune interferon that is produced primarily by T-lymphocytes and natural killer cells in response to antigens, mitogens, Staphylococcus enterotoxin B, phytohemaglutanin and other cytokines. IFN gamma is a dimeric protein consisting of two 146 amino acid subunits. IFN gamma is a glycoprotein that exists, functionally, as a homodimer of approximately 45 kDa. On SDS-PAGE, IFN gamma appears as a combination of 25, 20 and minor 15.5 kDa bands as a result of differential glycosylation. The biological activity of the IFN gamma homodimer is highly species specific. Human IFN gamma does not show cross-reactivity with mouse. IFN gamma function includes the following: antiviral activity, tumor antiproliferative activity, induction of class I and II MHC, macrophage activation, and enhanced immunoglobulin secretion by B lymphocytes. IFN gamma is involved in cytokine regulation and also acts synergistically with other cytokines. Activation of IFN gamma takes place through binding of IFN gamma receptor I and II, and activating the JAK-STAT pathway. IFN gamma does not show any homology with IFN alpha or IFN beta but human IFN gamma shows about 40% sequence homology with mouse IFN gamma. IFN gamma is upregulated by IL2, FGF basic, EGF and downregulated by vitamin D3 or DMN. IFN gamma gene mutations are associated with aplastic anemia.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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