IL-12/IL-35 p35 Antibody (27537) [Unconjugated]

PBMC's + LPS (1ug/mL;24 hrs) and monensin (3 hrs) (A) vs untreated PBMC's (B) were stained with Mouse Anti-Human/Mouse IL‑12/IL‑35 p35 Monoclonal Antibody (Catalog # mab1570) and Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). 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.

• Reactivity: Hu, Mu
• Applications: WB, CyTOF-ready, Flow
• Clone: 27537
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-12/IL-35 p35 Antibody (27537) [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑12/IL-35 p35
  Arg23-Ser219
  Accession # P29459
• 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: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: IL12A
• Purity Statement: Protein A or G purified from ascites

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-12/IL-35 p35 Antibody (27537) [Unconjugated]

• CLMF p35
• CLMF
• cytotoxic lymphocyte maturation factor 1, p35
• Cytotoxic lymphocyte maturation factor 35 kDa subunit
• IL-12, subunit p35
• IL-12/IL-35 p35
• IL-12A
• IL35 subunit
• IL35p35
• interleukin 12, p35
• interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocyte maturation factor 1, p35)
• interleukin-12 alpha chain
• interleukin-12 subunit alpha
• NF cell stimulatory factor chain 1
• NFSK
• NK cell stimulatory factor chain 1
• NKSF1
• p35

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-12/IL-35 p35 Antibody (MAB1570)(19)

Publications using MAB1570

• Nikolas T. Martin, Mathieu J. F. Crupi, Zaid Taha, Joanna Poutou, Jack T. Whelan, Sydney Vallati et al. Engineering Rapalog-Inducible Genetic Switches Based on Split-T7 Polymerase to Regulate Oncolytic Virus-Driven Production of Tumour-Localized IL-12 for Anti-Cancer Immunotherapy Pharmaceuticals (Basel) 2023-05-07 [PMID: 37242495] (Western Blot, Human, Mouse)
• Nikolas T. Martin, Mathieu J. F. Crupi, Zaid Taha, Joanna Poutou, Jack T. Whelan, Sydney Vallati et al. Engineering Rapalog-Inducible Genetic Switches Based on Split-T7 Polymerase to Regulate Oncolytic Virus-Driven Production of Tumour-Localized IL-12 for Anti-Cancer Immunotherapy Pharmaceuticals (Basel) 5/7/2023 [PMID: 37242495]
• Z Luo, C Soläng, R Larsson, K Singh Interleukin-35 Prevents the Elevation of the M1/M2 Ratio of Macrophages in Experimental Type 1 Diabetes International Journal of Molecular Sciences, 2022-07-19;23(14):. 2022-07-19 [PMID: 35887317] (Flow Cytometry, Mouse)
• A Wetzel, B Scholtka, F Schumacher, H Rawel, B Geisendörf, B Kleuser Epigenetic DNA Methylation of EBI3 Modulates Human Interleukin-35 Formation via NFkB Signaling: A Promising Therapeutic Option in Ulcerative Colitis International Journal of Molecular Sciences, 2021-05-19;22(10):. 2021-05-19 [PMID: 34069352] (Neutralization, Human)
• I Mizoguchi, M Ohashi, H Hasegawa, Y Chiba, N Orii, S Inoue, C Kawana, M Xu, K Sudo, K Fujita, M Kuroda, SI Hashimoto, K Matsushima, T Yoshimoto EBV-induced gene 3 augments IL-23R&alpha protein expression through a chaperone calnexin J. Clin. Invest., 2020-11-02;0(0):. 2020-11-02 [PMID: 32809973] (Western Blot, Mouse)
• Midhat H. Abdulreda, Dora M. Berman, Alexander Shishido, Christopher Martin, Maged Hossameldin, Ashley Tschiggfrie et al. Operational immune tolerance towards transplanted allogeneic pancreatic islets in mice and a non-human primate Diabetologia 5/1/2019 [PMID: 30701283]
• Jian Zhang, Tao Mao, Shuyun Wang, Dongsheng Wang, Zhaojian Niu, Zhenqing Sun et al. Interleukin-35 expression is associated with colon cancer progression Oncotarget 9/22/2017 [PMID: 29069729]
• Li Wang Gene expression and antiviral activity of interleukin-35 in response to influenza A virus infection J Biol Chem, 2016-06-15;0(0):. 2016-06-15 [PMID: 27307042] (Western Blot, Human)
• MM Halpert, V Konduri, D Liang, Y Chen, JB Wing, S Paust, JM Levitt, WK Decker Dendritic Cell-Secreted Cytotoxic T-Lymphocyte-Associated Protein-4 Regulates the T-cell Response by Downmodulating Bystander Surface B7 Stem Cells Dev., 2016-05-02;25(10):774-87. 2016-05-02 [PMID: 26979751] (Western Blot, Human)
• Liu G, Bi Y, Xue L, Zhang Y, Yang H, Chen X, Lu Y, Zhang Z, Liu H, Wang X, Wang R, Chu Y, Yang R Dendritic cell SIRT1-HIF1alpha axis programs the differentiation of CD4+ T cells through IL-12 and TGF-beta1. Proc Natl Acad Sci U S A, 2015-02-17;112(9):E957-65. 2015-02-17 [PMID: 25730867] (Bioassay, Mouse)
• Hsiao H, Hsu T, Liu W, Hsieh W, Chou T, Wu Y, Jiang S, Lai M Deltex1 antagonizes HIF-1alpha and sustains the stability of regulatory T cells in vivo. Nat Commun, 2015-02-19;6(0):6353. 2015-02-19 [PMID: 25695215] (Flow Cytometry, Mouse)
• Rossetti M, Spreafico R, Saidin S, Chua C, Moshref M, Leong J, Tan Y, Thumboo J, van Loosdregt J, Albani S Ex vivo-expanded but not in vitro-induced human regulatory T cells are candidates for cell therapy in autoimmune diseases thanks to stable demethylation of the FOXP3 regulatory T cell-specific demethylated region. J Immunol, 2014-12-01;194(1):113-24. 2014-12-01 [PMID: 25452562] (Neutralization, Human)
• YUGANG WANG, JINBIN DONG, WENYING MENG, JIALI MA, NA WANG, JUE WEI et al. Effects of phased joint intervention on IL-35 and IL-17 expression levels in patients with portal hypertension International Journal of Molecular Medicine 5/1/2014 [PMID: 24549402]
• Kochetkova I, Thornburg T, Callis G, Holderness K, Maddaloni M, Pascual D Oral Escherichia coli colonization factor antigen I fimbriae ameliorate arthritis via IL-35, not IL-27. J Immunol, 2013-12-11;192(2):804-16. 2013-12-11 [PMID: 24337375] (ELISA Development, Mouse)
• Olson B, Jankowska-Gan E, Becker J, Vignali D, Burlingham W, McNeel D Human prostate tumor antigen-specific CD8+ regulatory T cells are inhibited by CTLA-4 or IL-35 blockade. J Immunol, 2012-11-14;189(12):5590-601. 2012-11-14 [PMID: 23152566] (Neutralization, Human)
• Jindra C Human rhinoviruses induce IL-35-producing Treg via induction of B7-H1 (CD274) and sialoadhesin (CD169) on DC. Eur J Immunol, 2010-02-01;40(2):321-9. 2010-02-01 [PMID: 19950173] (Flow Cytometry, Immunoprecipitation, Human)
• Maneechotesuwan K, Wamanuttajinda V, Kasetsinsombat K, Huabprasert S, Yaikwawong M, Barnes PJ, Wongkajornsilp A Der p 1 suppresses indoleamine 2, 3-dioxygenase in dendritic cells from house dust mite-sensitive patients with asthma. J. Allergy Clin. Immunol., 2008-12-05;123(1):239-48. 2008-12-05 [PMID: 19058839] (Flow Cytometry, Human)
• Liu TY, Uemura Y, Suzuki M, Narita Y, Hirata S, Ohyama H, Ishihara O, Matsushita S Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells. Eur. J. Immunol., 2008-04-01;38(4):1012-23. 2008-04-01 [PMID: 18350544] (Flow Cytometry, Human)
• Nagarajan S, Selvaraj P Human tumor membrane vesicles modified to express glycolipid-anchored IL-12 by protein transfer induce T cell proliferation in vitro: a potential approach for local delivery of cytokines during vaccination. Vaccine, 2005-12-07;24(13):2264-74. 2005-12-07 [PMID: 16376465] (Western Blot, Human)

Bioinformatics

• Gene Symbol: IL12A
• Uniprot:
  • P29459()