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Key Product Details

Species Reactivity

Human

Applications

Immunocytochemistry, Intracellular Staining by Flow Cytometry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2038C

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL-12 p40 and human IL-12 p35
Ile23-Ser328 of p40, Arg23-Ser219 of p35
Accession # P29460 (p40) & P29459 (P35)

Specificity

Detects human IL-12/IL-35 p35 in direct ELISAs. Detects the IL-12/IL-35 p35 subunit either as part of the IL-12 p40/p35 heterodimer or as the IL-12/IL-35 p35 monomer. This antibody does not detect the IL-12/IL-23 p40 subunit.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human IL-12/IL-35 p35 Antibody

Detection of IL-12/IL-35 p35 antibody in Human PBMCs treated with rhIFN- ? and LPS antibody by Flow Cytometry.

Detection of IL‑12/IL‑35 p35 in Human PBMCs treated with rhIFN- gamma and LPS by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs), A) treated with recombinant human IFN- gamma (Catalog # 285-IF, 75 ng/mL for 2 hours), then LPS (1 µg/mL for 12 hours) and lastly monensin (3 µM for 3 hours), or B) untreated, were stained with Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (Catalog # FAB3832P) and Rabbit Anti-Human IL-12/IL-35 p35 Monoclonal Antibody (Catalog # MAB15701) followed by APC-conjugated Goat Anti-Rabbit Secondary Antibody (Catalog # F0111). Quadrant markers were set based on isotype control antibody (Catalog # MAB1050, data not shown). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
IL-12/IL-35 p35 antibody in Human PBMCs by Immunocytochemistry (ICC).

IL‑12/IL‑35 p35 in Human PBMCs.

IL-12/IL-35 p35 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Rabbit Anti-Human IL-12/IL-35 p35 Monoclonal Antibody (Catalog # MAB15701) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human IL-12/IL-35 p35 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs)

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Human PBMCs treated with rhIFN- gamma and LPS, fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in 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, 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. Current 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 p35

Alternate Names

IL-12A, IL35p35

Entrez Gene IDs

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

Gene Symbol

IL12A

Additional IL-12/IL-35 p35 Products

Product Documents for Human IL-12/IL-35 p35 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-12/IL-35 p35 Antibody

For research use only

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