Human GDF-15 ELISA Kit

Principle of the Assay

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for GDF-15 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any GDF-15 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for GDF-15 is added to the wells and binds to the combination of capture antibody- GDF-15 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 GDF-15 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 GDF-15 standard dilutions and GDF-15 sample concentration determined.

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

Target Information

Growth and differentiation factor-15 (GDF-15), also known as macrophage inhibitory cytokine-1(MIC-1), placental transforming growth factor-β, prostate-derived factor, and placental bone morphogenetic protein, is a divergent member of the transforming growth factor beta (TGF-β) superfamily. GDF-15 is initially synthesized as a 40 kDa inactive precursor protein. It is then proteolytically cleaved to release the active C-terminal fragment, which is assembled into a disulfide-linked homodimer of 28 kDa to become biologically active GDF-15. In humans, GDF-15 is predominantly expressed in the placenta, with low levels in the kidney, pancreas, and prostate. However, its expression can be rapidly induced by cytokines such as interleukin-1 and TGF-β.

GDF-15 has diverse biological functions. Early studies have shown that low serum GDF-15 levels correlate with miscarriages, indicating that it might be able to suppress inflammation in early pregnancy. GDF-15 also plays an important role in tumorigenesis and metastasis. It has been observed that in many types of cancers, such as colorectal, breast, and prostate, the expression of GDF-15 is dramatically increased. Additionally, in cancer patients, serum levels of GDF-15 are elevated, which are of value in disease diagnosis and stratification. GDF-15 is strongly induced by the tumor suppressor gene p53 and other anti-tumorigenic agents, such as the non-steroidal anti-inflammatory drugs and peroxisome proliferators-activated receptor γ. These findings suggest that GDF-15 may be a downstream target of those signaling pathways that regulate cell cycle arrest and apoptosis. Through the modulation of neuronal pathways important in the regulation of appetite and energy homeostasis, GDF-15 mediates cancer-induced anorexia and weight loss.

GENE ID 9518
SWISS PROT Q99988
SYNONYMS PDF; MIC1; PLAB; MIC-1; NAG-1; PTGFB; GDF-15

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human GDF-15 (8﹡12) 1 plate
Human GDF-15 Standard lyophilized, 1000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human GDF-15 monoclonal antibody 2 vials
Streptavidin-HRP solution 2 vials
Standard /sample Diluent 1 bottle
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 GDF-15. The factors listed below were prepared at 50ng/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 GDF-11, GDF-5, GDF-6, GDF-7, GDF-8, EGF, CSF, sTNF-RII, FGF, TNF-α, PDGF
Recombinant mouse GDF-11, GDF-5, GDF-6, GDF-7, GDF-8, GDF-9