Human IL-2 Enzyme Immunoassay (EIA)

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Tony Rook
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Human IL-2 Enzyme Immunoassay (EIA)

Please find the link for the following protocol:

Human IL-2 Enzyme Immunoassay (EIA)

Reference Link:
http://aactg.s-3.com/pub/download/imm/actgil2b.pdf

PRINCIPLE AND PURPOSE OF TEST

Interleukin-2 (IL-2) is a multi-functional protein most notably known for the ability to induce the proliferation and maturation of activated T cells. Additional biological
functions include:

Stimulus for the division of activated natural killer cells and tumor-infiltrating lymphocytes plus the enhancement of the ability of theses cytotoxic lymphocytes to kill targets cells

Stimulus for the division of antibody-producing B cells

Up regulation of IL-2 receptors on T cells

Increased division and maturation of activated helper T cells

Stimulation of neutrophils

Stimulus for the secretion of interferon-( and tumor necrosis factors - aphla and - Beta from peripheral mononuclear cells.

The Endogen Human IL-2 EIA is an in vitro enzyme-linked immunosorbent assay for quantitatively detecting human IL-2 in biological fluids (e.g. serum, plasma) and in cell culture supernatants. The kit consists of a 96 well microtiter plate that has been precoated with anti-IL-2 antibodies for the capture of human IL-2 from specimens. An IL-2 standard is provided and is simultaneously processed with specimens. In order to detect
the captured IL-2 a second anti-IL-2 antibody that has been biotinylated is added to all wells. This results in a sandwiching of any IL-2 (capture IL-2 Ab C IL-2 C detection IL-2- biotin Ab). Following the removal of unbound antibodies, by a series of washings, a horseradish peroxidase (HRP) conjugate with a high affinity for biotin is added. Unbound Strepavidin-HRP is removed and the bound enzyme labeled antibodies can then be measured via a chromogenic reaction with the addition of TMB substrate. The resulting chromogenic reaction is stopped using H SO (Stop Solution) and the optical 2 4 density of each well is then read at the appropriate wavelength. The level of substrate conversion is colormeterically determined by measuring the absorbance that is proportional to the amount of IL-2. The IL-2 standard is plotted and the cytokine concentration in the specimens is determined.