Thermo Fisher Scientific IL-12 p35 Monoclonal Antibody (4D10p35), eFluor 660, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.125 µg/test

View 3 publications 3 publications

ELISA (ELISA)

-

View 1 publication 1 publication

Product Specifications

Species Reactivity

Mouse

Published species

Human, Mouse

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), eFluor™ 660, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

4D10p35

Immunogen

Mouse IL-35-Fc recombinant protein

Conjugate

eFluor™ 660 eFluor™ 660 eFluor™ 660

Excitation/Emission Max

651/668 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2574284

Product Specific Information

Description: This 4D10p35 monoclonal antibody reacts with mouse p35, a component of two of the four members of the interleukin (IL)-12 family of cytokines. The IL-12 family members are heterodimeric cytokines composed of an alpha-chain, such as p35, and a beta-chain, such as p40. The alpha-chain proteins are thought to be the limiting factors for secretion of IL-12 family member cytokines, as expression of the alpha-chain is more restricted than that of the beta-chain proteins, and the two subunits may need to be co-expressed in order for cytokine secretion to occur. IL-12 family members are secreted primarily by antigen-presenting cells in response to various TLR agonists and other inflammatory stimuli. The exact types of stimuli present and kinetics of stimulation determine the balance of IL-12 family member production.

The p35 protein was first identified as part of the IL-12 cytokine, which is also known as IL-12p70. Secretion is induced by IFN gamma and agonists of TLR3, 4, or 7 and is inhibited by Th2 cytokines. IL-12 is essential for the differentiation and maintenance of Th1 cells. The other subunit, p40, is expressed in much higher quantities than p35, and may be secreted independently as a monomer or homodimer. Free p40 may act as an antagonist of IL-12 function.

More recently, p35 was also found to dimerize with EBI3 (Epstein-Barr Virus-Induced Gene 3) to form IL-35. The biology of IL-35 is incompletely understood, and it is believed to be unique among the IL-12 family members in its expression and function. It is produced by regulatory T cells, and appears to have immunoregulatory functions. IL-35 has been found to suppress the proliferation and function of effector T cells, and has also been observed to induce a regulatory phenotype on naive CD4+ cells cultured in its presence.

Testing of 4D10p35 in the context of IL-35 has not been evaluated. For additional information, please contact technical support.

Applications Reported: This 4D10p35 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 4D10p35 antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse dendritic cells. This can be used at less than or equal to 0.125 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.

Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Interleukin 12A is a novel signaling cytokine belonging to the Interleukin-6 protein superfamily. Interleukin-12 alpha chain precursor (IL-12A) is a 35-kD subunit of disulfide-linked heterodimer of cytokine IL-12 and has a broad array of biological activities. Reports suggest that IL-12A can act as a growth factor for activated T and NK cells and enhance the lytic activity of NK/lymphokine-activated Killer cells. IL-12A stimulates the production of IFN-gamma by resting PBMC as well as T-cell-independent induction of interferon (IFN)-gamma, and helps in the differentiation of both Th1 and Th2 cells. It is also involved in positive regulation of tyrosine phosphorylation of Stat4 protein. The signaling process of IL-12A in innate immunity requires nitric oxide synthase 2A. Along with EBI3, IL-12A forms a novel heterodimeric cytokine, designated as interleukin-35 (IL-35), that may be specifically produced by T reg cells and is required for maximal suppressive activity.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.