Immunoassays | Inflammation & Cytokines

Erenna® IL-12 Immunoassay Kit (Cat# 03-0057-xx)

download PDF data sheet

download PDF package insert

Introduction

The Erenna® IL-12 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IL-12 in human serum and plasma samples. A capture antibody specific for human IL-12 has been pre-coated onto paramagnetic microparticles (MPs). The user pipettes MPs, standards, and samples into uncoated microplate wells. During incubation, the IL-12 present in the sample binds to the capture antibody on the coated MPs. Unbound molecules are washed away during the subsequent buffer exchange and wash steps. Fluor-labeled detection antibody is added to each well and incubated. This detection antibody recognizes and binds to IL-12 that has been captured onto the MPs. During the following wash step the MPs are transferred to a clean plate. Elution buffer is then added and incubated. The elution buffer dissociates the bound protein sandwich from the MP surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final microplate. The plate is loaded into the Erenna® System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IL-12 present in the sample when captured. The amount of IL-12 in unknown samples is interpolated from a standard curve.

Assay Performance

¹ See product insert for updated values
² LLoQ = 20% CV and ± 20% recovery
³ based upon median [IL-12] in a healthy reference

FIGURE 1. [IL-12] in EDTA plasma from 10 healthy donors, with median and interquartile range. The Erenna® IL-12 Immunoassay Evaluation Reagent Kit reliably quantifies IL-12 in healthy subjects, who have a median [IL-12] of 0.13 pg/mL that is well above the detection limit of 0.01 pg/mL.

FIGURE 2. Erenna® IL-12 Immunoassay Evaluation Reagent Kit low-end standard curve signal (left) and curve fit (right).

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Interleukin-12 (IL-12) is a cytokine expressed by T-cells and natural killer cells in response to the presence of microbial agents. Increased expression of IL-12 drives a positive feedback loop, which generates more T-cells and natural killer cells, along with other immuno-regulatory cytokines such as IFN-γ, IL-10 and TNFα. Although IL-12 is an important initiator of cell-mediated immunity, its pro-inflammatory effects can be deleterious. Over production of this cytokine has been implicated in several autoimmune disorders such as inflammatory bowel disease, psoriasis, and Crohn’s disease. The ability to escalate and strengthen the immune response makes IL-12 an important immunostimulant therapy for infectious diseases and metastatic cancers.

* Source Info from www.uniprot.org.

Related Products