Human Resistin ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for Resistin has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Resistin present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for Resistin is added to the wells and binds to the combination of capture antibody- Resistin in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of Resistin present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven Resistin standard dilutions and Resistin sample concentration determined.

For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Resistin, also known as Found In Inflammatory Zone 3 (FIZZ3) or Adipocyte Secreted Factor (ADSF), is a member of a protein family known as the Resistin-like molecules (RELMs). It is perhaps best known for its potential as a link between obesity and the development of insulin resistance. Other members of this family include RELM-α/FIZZ1 and RELM-γ, which are described in rodents but as yet have no identified human counterparts, and RELM-β/FIZZ2. The Resistin amino acid (aa) sequence contains a putative N-terminal signal sequence and a motif containing 11 cysteine residues, 10 of which are characteristic of the RELM family. The protein is thought to be secreted as a dimer and potentially exists in higher order molecular structures resulting from interactions between Resistin dimers or other members of the RELM family. A splice variant in the rat, lacking the signal sequence and localized predominantly to the nucleus, has also been described. A large 3' intron is the primary reason that the mouse genomic sequence is 3-fold larger than the corresponding human sequence. Mouse and human Resistin share only 53 percent identity at the aa level and exhibit differences in expression patterns. In mouse, expression appears primarily in adipose tissues. Although some human studies suggest Resistin is expressed by adipose tissues as well, the most significant source appears to be blood mononuclear cells. In humans, Resistin is also reported to be expressed by pre-adipocytes, placenta, pancreatic islets, and primary leukemia cells. A receptor for Resistin has not yet been described.

Resistin acquired initial attention as a potential link between obesity and glucose regulation. Serum levels were shown to increase in diet-induced and genetic forms of obesity in mice (ob/ob and db/db) and decrease in response to insulin sensitizing drugs (TZDs). In addition, function-blocking Resistin antibodies enhanced insulin actions while treatment with recombinant Resistin caused glucose intolerance and insulin resistance. Resistin knockout mice exhibit decreased fasting blood glucose levels as a result of reduced hepatic output. To establish a physiological role in humans, several studies have examined whether altered circulating Resistin levels are associated with type 2 diabetes, insulin resistance, and/or obesity. Although some demonstrate significant correlations, others report no correlation, suggesting that in humans fundamental questions remain regarding Resistin’s role in these pathophysiological processes. Resistin expression by human mononuclear cells could indicate a potential role in inflammation. In vitro, Resistin expression by these cells is enhanced by treatment with several pro-inflammatory cytokines including IL-1β, TNF-α, IFN-γ, or IL-6. In addition, Resistin has been shown to activate endothelial cells in vitro, leading to the production of adhesion molecules, Endothelin-1, and chemokines.

GENE ID 56729
SWISS PROT Q9HD89
SYNONYMS ADSF; RSTN; XCP1; FIZZ3; RETN1

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human Resistin (8﹡12) 1 plate
Human Resistin Standard lyophilized, 1000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human Resistin monoclonal antibody 2 vials
Streptavidin-HRP solution 2 vials
Standard /sample Diluent 4 bottles
Biotin-Conjugate antibody Diluent 1 bottle
Streptavidin-HRP Diluent 1 bottle
Wash Buffer Concentrate 20x (PBS with 1% Tween-20) 1 bottle
Substrate Solution 1 vial
Stop Solution 1 vial
Adhesive Films 4 pieces
Product data sheet 1 copy

Storage

Storage Store at 2 - 8°C

Performance Characteristics

REPEATABILITY The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY The minimum detectable dose was 4pg/mL.
ASSAY RANGE 15.6 - 1000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant human Resistin. The factors listed below were prepared at 20ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.
Factors assayed for cross-reactivity
Recombinant human G-CSF, GM-CSF, IL-1α, IL-1β, IL-2 sRα
Recombinant mouse IL-1α, IL-1β, IL-3, IL-5, IL-6