IL-10 Monoclonal Antibody (JES3-9D7), PerCP-eFluor™ 710, eBioscience™

Product Specific Information

Description: The JES3-9D7 monoclonal antibody reacts with human interleukin-10 (IL-10).

Applications Reported: This JES3-9D7 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This JES3-9D7 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. This can be used at 5 µL (0.03 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.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

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 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

IL-10 encodes a protein that acts as a cytokine and is primarily produced by monocytes, with some production by lymphocytes. This cytokine has various effects on immunoregulation and inflammation, including down-regulating the expression of Th1 cytokines, MHC class II Ags, and costimulatory molecules on macrophages. It also enhances B cell survival, proliferation, and antibody production. Additionally, IL-10 can block NF-kappa B activity and is involved in the regulation of the JAK-STAT signaling pathway. Knockout studies in mice have suggested that this cytokine is an essential immunoregulator in the intestinal tract. Mutations in this gene have been associated with an increased susceptibility to HIV-1 infection and rheumatoid arthritis. Diseases associated with IL10 include Graft-Versus-Host Disease and Human Immunodeficiency Virus Type 1.