sHLA-G ELISA
HLA-G differs from the other MHC class I genes by its low polymorphism and alternative splicing that generates seven HLA-G proteins, whose tissue-distribution is restricted to normal fetal and adult tissues that display a tolerogeneic function toward both innate and acquired immune cells. Soluble HLA-G is an immunosuppressive molecule inducing apoptosis of activated CD8(+) T cells and down-modulating CD4(+) T cell proliferation. Recently, using specific ELISA to analyse the presence of sHLA-G molecules in culture supernatants of early embryos obtained by in vitro fertilization (IVF) before transfer, several reports demonstrated that positive embryo implantations occurred with embryo secreting sHLA-G molecules. These breakthrough results indicate that sHLA-G ELISA can be a useful biochemical assay in addition to embryo morphology in embryo selection for transfer in IVF treatment if there are other embryos with the same morphology. Furthermore, monitoring of sHLA-G in amniotic fluid and plasma of pregnant women may have an important prognostic value to recognize pathological situations. Other interesting observations suggest that HLA-G molecules seem to be directly involved in transplant acceptation, and their analysis should be taken into consideration when monitoring transplant-patients status. In addition, soluble HLA-G plasma levels are increased in lyphoproliferative disorders or in patients suffering from malignant melanoma, glioma, breast and ovarian cancer.
The RD194070100R sHLA-G ELISA is a sandwich enzyme immunoassay for the quantitative measurement of soluble forms of human leukocyte antigen-G (sHLA-G).
In the BioVendor sHLA-G ELISA, calibrators and samples are incubated in microplate wells pre-coated with monoclonal anti-sHLA-G antibody. After 16-20 hours incubation and washing, monoclonal anti-human β2-microglobulin antibody labelled with horseradish peroxidase (HRP) is added to the wells and incubated for 60 minutes with captured sHLA-G. Following another washing step, the remaining HRP conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured. The absorbance is proportional to the concentration of sHLA-G. A calibration curve is constructed by plotting absorbance values against concentrations of calibrators, and concentrations of unknown samples are determined using this calibration curve.
Intended use
Clinical Application
Test principle
Summary of protocol
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