Human IL-2 Quantikine QuicKit ELISA Summary
Cell Culture Supernates, Serum, Plasma, EDTA Plasma, Heparin Plasma
|Intra-Assay Precision||Inter-Assay Precision|
The recovery of human IL-2 spiked to three levels in samples throughout the range of the assay was evaluated.
|Sample Type||Average % Recovery||Range %|
|Cell Culture Media (n=4)||112||103-119|
|EDTA Plasma (n=2)||85||72-96|
|Heparin Plasma (n=2)||85||74-93|
Human IL-2 ELISA Standard Curve
Human IL-2 QuicKit Spiked Recovery Competitor Comparison IL-2 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 103% compared to 113% for the top competitor. EDTA Plasma recovery is 107% compared to 111% for the top competitor. Heparin Plasma recovery is 100% compared to 102% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.
Preparation and Storage
Interleukin 2 (IL-2), also known as T cell growth factor (TCGF), is a 15-18 kDa variably glycosylated alpha -helical polypeptide that is a member of the Common gamma Chain ( gamma c) cytokine family (1-4). It exists as a monomer and has a notably short half-life (< 30 minutes) (1). Human IL-2 is synthesized as a 153 amino acid (aa) precursor that contains a 20 aa signal sequence plus a 133 aa mature region (5, 6). The mature region is alpha -helical in nature, and contains one utilized O-linked glycosylation site at Thr3 plus three cysteines, two of which form an intrachain disulfide bond that is essential for activity (7). Mature human IL-2 shares 73%, 66%, 78% and 97% aa identity with canine, rat, feline and rhesus monkey IL-2, respectively. Although human IL-2 shares only approximately 60% aa identity with the highly polymorphic mouse IL-2, human IL-2 is known to be active on mouse IL-2 responsive cells. Cells reported to secrete IL-2 include gamma δ T cells (8), activated conventional CD4+ and CD8+ T cells (1, 9), neurons (10, 11), microglia (12), and hematopoietic stem cells (13).
These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.
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