• Species Reactivity

  Human, Mouse

• Specificity

  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.

• Source

  Monoclonal Mouse IgG    ₁ Clone # 27537

• Purification

  Protein A or G purified from ascites

• Immunogen

  S. frugiperda insect ovarian cell line     Sf 21-derived recombinant human IL‑12/IL-35 p35    
  Arg23-Ser219    
  Accession # P29459

• Formulation

  Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

• Label

  Unconjugated

Applications

• Recommended    
  Concentration

  Sample

• Western Blot

  1 µg/mL

  Recombinant Human IL-12 (Catalog # )

• 
  

• CyTOF-ready

  Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

• 
  

• Intracellular Staining by Flow Cytometry

  2.5 µg/10    ⁶ cells

  Human peripheral blood mononuclear cells or mouse splenocytes treated with LPS, fixed with paraformaldehyde, and permeabilized with saponin

• 
  

Please Note: Optimal dilutions should be determined by each laboratory for each application.  are available in the Technical Information section on our website.

Preparation and Storage

• Reconstitution

  Reconstitute at 0.5 mg/mL in sterile PBS.

• Shipping

  The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

• Stability & Storage

  Use a manual defrost freezer and avoid repeated freeze-thaw cycles.    

• 12 months from date of receipt, -20 to -70 °C as supplied.

• 1 month, 2 to 8 °C under sterile conditions after reconstitution.

• 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-12/IL-35 p35

Interleukin 12 (IL-12) and interleukin 35 (IL-35) are heterodimeric cytokines composed of alpha and beta chains. IL-12 is composed of p35 and p40 subunits, while IL-35 is comprised of p35 paired with EBI-3 (1). In mice, IL-35 is produced by FoxP3  ⁺ regulatory T cells and may function as an inhibitory cytokine to suppress T cell proliferation (2). Human FoxP3  ⁺ Tregs do not constitutively express IL-35 (3), but expression may be induced by activated dendritic cells (4).

• References:

1. Collison, L.W. and D.A.A. Vignali (2008) Immunol. Rev. 226:248.

2. Collison, L.W. et al. (2007) Nature 450:566.

3. Bardel, E. et al. (2008) J. Immunol. 181:6898.

4. Seyerl, M. et al.(2010) Eur. J. Immunol. 40:321.

• Long Name:

  Interleukin 12 p35

• Entrez Gene IDs:

  3592 (Human); 16159 (Mouse); 84405 (Rat)

• Alternate Names:

  IL-12/IL-35 p35; IL-12A; IL35p35