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 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.
Human IL-2 ELISA Standard Curve
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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