Human IL-12 Biotinylated Antibody

Catalog #: BAF219
13 Citations Datasheet / COA / SDS

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BAF219

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Citations (13)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human IL-12 Biotinylated Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL-12 in ELISAs and Western blots. When used in combination with the monoclonal capture antibody anti‑human IL-12 p40 (Catalog # MAB609) in sandwich ELISAs, less than 2.5% cross‑reactivity was observed with rhIL-12 p70. When used in combination with the monoclonal capture antibody anti-human IL-12 p70 (Catalog # MAB611) in sandwich ELISAs, less than 0.1% cross‑reactivity was observed with rhIL‑12 p40, rhIL‑12 p35 and rmIL‑12.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑12 heterodimer
Arg23-Ser219 of p35; Ile23-Ser328 of p40
Accession # P29459 (p35) & P29460 (p40)

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human IL‑12 (Catalog # 219-IL)

Immunocytochemistry

5-15 µg/mL

See below

Human IL-12 p70 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Detection (Matched Antibody Pair)

0.1-0.4 µg/mL

Use in combination with:

Capture Reagent: Human IL‑12 p70 Antibody (Catalog # MAB611)

Capture Reagent: Human IL‑12 p70 Antibody (Catalog # MAB611R)

Standard: Recombinant Human IL-12 Protein (Catalog # 219-IL)

Human IL-12/IL-23 p40 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Detection (Matched Antibody Pair)

0.1-0.4 µg/mL

Use in combination with:

Capture Reagent: Human IL‑12/IL‑23 p40 Antibody (Catalog # MAB609)

Standard: Recombinant Human IL-12/IL-23 p40 Monomer Protein (Catalog # 309-IL)

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.

Scientific Data

Immunocytochemistry

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IL‑12 in Human PBMCs. IL-12 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with calcium ionomycin and PMA using Goat Anti-Human IL-12 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF219) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

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

Interleukin 12, also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a pleiotropic cytokine originally identified in the medium of activated human B lymphoblastoid cell lines. The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the human IL-6 receptor while the p35 subunit shows distant but significant sequence similarity to IL-6, G-CSF, and chicken MGF. These observations have led to the suggestion that IL-12 might have evolved from a cytokine/soluble receptor complex. Human and murine IL-12 share 70% and 60% amino acid sequence homology in their p40 and p35 subunits, respectively. IL-12 apparently shows species specificity with human IL-12 reportedly showing minimal activity in the murine system.

IL-12 is produced by macrophages and B lymphocytes and has been shown to have multiple effects on T cells and natural killer (NK) cells. These effects include inducing production of IFN-gamma and TNF by resting and activated T and NK cells, synergizing with other IFN-gamma inducers at both the transcriptional and post-transcriptional levels. This interaction induces IFN-gamma gene expression, enhancing the cytotoxic activity of resting NK and T cells, inducing and synergizing with IL-2 in the generation of lymphokine-activated killer (LAK) cells, acting as a co-mitogen to stimulate proliferation of resting T cells, and inducing proliferation of activated T and NK cells. Evidence indicates that IL-12, produced by macrophages in response to infectious agents, is a central mediator of the cell-mediated immune response by its actions on the development, proliferation, and activities of TH1 cells. In its role as the initiator of cell-mediated immunity, it has been suggested that IL-12 has therapeutic potential as a stimulator of cell-mediated immune responses to microbial pathogens, metastatic cancers, and viral infections such as AIDS.

Long Name

Interleukin 12

Entrez Gene IDs

3592 (Human); 16159 (Mouse); 84405 (Rat); 397053 (Porcine); 403977 (Canine); 102115316 (Cynomolgus Monkey); 493741 (Feline); 703205 (Rhesus Macaque)

Alternate Names

CLMF p35; CLMF1; Cytotoxic lymphocyte maturation factor 35 kDa subunit; IL-12 subunit p35; IL12; IL-12; IL-12, subunit p35; IL12A; interleukin 12, p35; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35); interleukin-12 alpha chain; interleukin-12 subunit alpha; natural killer cell stimulatory factor 1, 35 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NKSF1; p35

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human IL-12 Biotinylated Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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A Gel-Based Dual Antibody Capture and Detection Method for Assaying of Extracellular Cytokine Secretion: EliCell
Authors: Lisa A. Spencer, Rossana C. N. Melo, Sandra A. C. Perez, Peter F. Weller
Handbook of ELISPOT
mWTX-330, an IL 12 INDUKINE Molecule, Activates and Reshapes Tumor-infiltrating CD8+ T and NK Cells to Generate Antitumor Immunity
Authors: CJ Nirschl, HR Brodkin, C Domonkos, CJ Dwyer, DJ Hicklin, N Ismail, C Seidel-Dug, P Steiner, Z Steuert, JM Sullivan, WM Winston, A Salmeron
Cancer Immunology Research, 2023-04-19;0(0):.
Species: Human
Sample Types: Plasma, Tissue Homogenates
Applications: ELISA Detection
Cell surface-tethered IL-12 repolarizes the tumor immune microenvironment to enhance the efficacy of adoptive T cell therapy
Authors: DS Jones, JD Nardozzi, KL Sackton, G Ahmad, E Christense, L Ringgaard, DK Chang, DE Jaehger, JV Konakondla, M Wiinberg, KL Stokes, A Pratama, K Sauer, TL Andresen
Science Advances, 2022-04-27;8(17):eabi8075.
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Detection
Tryptophan (W) at position 37 of murine IL-12/IL-23 p40 is mandatory for binding to IL-12Rbeta1 and subsequent signal transduction
Authors: J Georgy, Y Arlt, JM Moll, M Ouzin, HT Weitz, L Gremer, D Willbold, J Grötzinger, F Thives-Kur, J Scheller, DM Floss
The Journal of Biological Chemistry, 2021-10-09;0(0):101295.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot
Preclinical development of a bispecific TNFalpha/IL-23 neutralising domain antibody as a novel oral treatment for inflammatory bowel disease
Authors: KJ Roberts, MF Cubitt, TM Carlton, L Rodrigues-, L Maggiore, R Chai, S Clare, K Harcourt, TT MacDonald, KP Ray, A Vossenkämp, MR West, JS Crowe
Scientific Reports, 2021-09-30;11(1):19422.
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Detection
TLR-stimulated CD34 stem cell-derived human skin-like and monocyte-derived dendritic cells fail to induce Th17 polarization of naive T cells but do stimulate Th1 and Th17 memory responses.
Authors: Duraisingham SS, Hornig J, Gotch F, Patterson S
J. Immunol., 2009-07-22;183(4):2242-51.
Species: Human
Sample Types: Whole Cells
Applications: ELISA Development
Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo.
Authors: Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB
AIDS Res. Hum. Retroviruses, 2008-03-01;24(3):453-62.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Luminex Development
Effect of serum content and diluent selection on assay sensitivity and signal intensity in multiplex bead-based immunoassays.
Authors: Pfleger C, Schloot N, ter Veld F
J. Immunol. Methods, 2007-10-22;329(1):214-8.
Species: Human
Sample Types: Serum
Applications: Luminex Development
Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1.
Authors: Biancotto A, Grivel JC, Iglehart SJ, Vanpouille C, Lisco A, Sieg SF, Debernardo R, Garate K, Rodriguez B, Margolis LB, Lederman MM
Blood, 2007-02-08;109(10):4272-9.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Luminex Development
HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1.
Authors: Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, Shattock RJ, McGowan I, Margolis L, Anton P
AIDS, 2007;21(10):1263-72.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Luminex Development

Blockade of TLR9 agonist-induced type I interferons promotes inflammatory cytokine IFN-gamma and IL-17 secretion by activated human PBMC.
Authors: Meyers JA, Mangini AJ, Nagai T, Roff CF, Sehy D, van Seventer JM
Cytokine, 2006-10-18;35(5):235-46.
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Development

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.
Authors: Nagarajan S, Selvaraj P
Vaccine, 2005-12-07;24(13):2264-74.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines.
Authors: Copeland S, Siddiqui J, Remick D
J. Immunol. Methods, 2004-01-01;284(1):99-106.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Array Development