Detects human IL‑12/IL-35 p35 in direct ELISAs and Western blots. Detects the p35 subunit either as part of a p40/p35 heterodimer or as a free subunit after reduction of the heterodimer. This antibody does not recognize IL-12 p40 homodimers but shows strong cross-reactivity with the p35 subunits from porcine and mouse systems.
Monoclonal Mouse IgG1 Clone # 27537
Protein A or G purified from hybridoma culture supernatant
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑12/IL-35 p35 Arg23-Ser219 Accession # P29459
Supplied in a saline solution containing BSA and Sodium Azide.
Intracellular Staining by Flow Cytometry
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of IL‑12/IL‑35 p35 in Human PBMCs or Mouse Splenocytes by Flow Cytometry. (A) Human peripheral blood mononuclear cell (PBMCs) or (B) mouse splenocytes either untreated (light orange filled histogram) or treated with LPS (dark orange filled histogram) were stained with Mouse Anti-Human/Mouse IL‑12/IL‑35 p35 APC‑conjugated Monoclonal Antibody (Catalog # IC2191A) or isotype control antibody (Catalog # IC002A, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
Background: IL-12/IL-35 p35
is a 34-36 kDa monomeric member of the IL-6 superfamily of molecules. It is 219 amino acids in length, and possibly
contains an N-terminal 22 aa signal sequence.
In any event, it does not appear to be individually secreted, and its
release from cells is commonly assumed to be as part of a disulfide-linked
heterodimer linked to either p40 (forming IL-12) or EBI3 (forming IL-35). Multiple cell types are known to express p35,
and include keratinocytes, CD138+ plasma cells, naïve B cells, monocyte-derived
dendritic cells, macrophages, CD86lo B7-H1hi CD206+
B7-DC+ tolerogenic dendritic cells, neutrophils, activated monocytes and
macrophages, trophoblasts, testicular endothelial cells, and T cells, including
CD4+ and CD8+ T helper cells, and CD4+ CD25+ Tregs. The presence of the subunit does not
necessarily indicate function, as the p35:p40 heterodimer is associated with
promoting inflammatory IFN-gamma production by NK cells, while the p35:EBI3
heterodimer is anti-inflammatory through its inhibition of Th17 cell
development. Over aa 23-219, human p35
shares 83% and 57% aa sequence identity with pig and mouse p35, respectively.
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