High Sensitive Resistin ELISA

$1,030.00

The Eagle Biosciences Resistin ELISA Assay Kit is intended  for the quantitative determination of human Resistin in serum, plasma, cell culture supernatants, breast milk, saliva and urine. The Eagle Biosciences Resistin ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80885 Categories: ,

High Sensitive Resistin ELISA

The High Sensitive Resistin ELISA is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.006 ng/mL
Dynamic Range: 0.02-1 ng/mL
Incubation Time: 5 hours
Sample Type: Serum, Plasma, cell culture supernatants, breast milk, saliva, urine
Sample Size: 100 µL


Assay Background

Resistin, a cysteine-rich protein of 11.3 kDa, was firstly found in mice and constitutes together with RELMα, RELMβ and RELMγ the protein family of resistin-like molecules (RELM). In humans, resistin and RELMβ but no other proteins of the RELM family were found. The human form of resistin shows a homology of 53% to the murine protein. It has 11 cysteine-residues, is synthesized as a propetide of 108 amino acids and secreted as a dimer; build by a disulfide bridge of cysteine residues. Beside this intermolecular disulfide bridge, 5 additional intramolecular ones exist. Appearance of multi- and oligomer formation was proved by size exclusion chromatography. Thereby it was shown, that oligomer formation is SDS-insensitive but can be inhibited by β-mercaptoethanol and is therefore likely to be caused by disulfide bridges. Further on, the resistin structure seems to be dependent on its concentration, as circular dichroism analysis shows a concentration dependent shift of αhelical to β-sheet structure. Resistin expression was demonstrated in white adipose tissue, pituitary and pancreatic islets of mice as well as in brown adipose tissue of rats. In humans, resistin expression in adipocytes can be detected but only at a very low level. But in vitro, resistin expression of nonadipocytes in fatty tissue was shown. Human resistin gene is also expressed in pancreatic islets, pre-adipocytes marcophages and bone marrow. So, resistin is of relevance for inflammation processes as well as for lipid metabolism. Most investigation refers to the mouse model. Here, the existence of trimeric and hexameric resistin in serum was demonstrated. In comparison to adiponectin biology it is highly probable that different resistin oligomers have different biologic function. In mice, a correlation between adiposity, insulin resistance and resistin expression was found empirically. In humans, respective study results are not clear – several studies show an association of resistin serum concentration and adiposity or insulin resistance. But others failed in confirming these results. Therefore, there is requirement for valid and reproducible determination of resistin serum concentration. Relevance of resistin in other physiologic processes than energy metabolism was investigated by several different approaches. Experiments with endothelial cells gave interesting results. Here, resistin was shown to enhance expression of VCAM-1 and ICAM-1. By this way, resistin is potentially able to influence endothelial inflammation and, thereby atherosclerosis. These results were confirmed by experiments in mice, where endothelin-1 was shown to regulate resistin secretion. In recent research human resistin was shown to increase pre-adipocyte proliferation and lipolysis of mature adipocytes. By the way of modulating MAPK-signalling pathways resistin exerts crucial influence on energy metabolism. Present research demonstrates, that Resistin exerts influence on a broad variety of physiological processes, however a clear and defined biological role of resistin remains still unexisting. This ELISA-kit enables the user to determine the exact concentration of Resistin in human serum/plasma as well as other body fluids and thereby assists investigation of Resistin biology.


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Additional Information

Assay Principle


This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Resistin has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Resistin present is bound by the immobilized antibody. After washing away any unbound substances, a Biotin-Resistin antibody conjugate is added to each well and incubate. After washing, HRP-Streptavidin is added and incubate. Then washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Renin bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450 nm ±2 nm. The concentration of Resistin in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure


1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
2. Add 100 µl Sample Buffer into blank wells.
3. Add 100 µl standards, controls and samples into each well.
4. Cover wells and incubate for 2 hours at RT.
5. Aspirate each well and wash, repeating the process 4 times for a total 5 washes. Wash by filling each well with wash buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining distilled water by aspirating, decanting or blotting against clean paper towels.
6. Add 100 µl Biotin-antibody conjugate into each well. Incubate for 60 minutes at RT.
7. Aspirate and wash wells as step 5.
8. Add 100 μl of HRP-Streptavidin conjugate into each well. Incubate for 30 minutes at RT.
9. Aspirate and wash wells as step 5.
10. Add 100 μl of TMB Substrate to each well. Incubate for 30 minutes at room temperature in dark.
11. Add 100 μl Stop solution into each well.
12. Read the OD with a microplate reader at 450 nm immediately.

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