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enzyme immunoassay kit,
enzyme assay kits
Human Customized GPR173 ELISA Test Kit Probable G-protein Coupled Receptor 173 ELISA Assay Kit
Standard Curve Range: 20ng/L - 6000ng/L
Size: 96 wells
This sandwich kit is for the accurate quantitative detection of Human Probable G-protein Coupled Receptor 173 (also known as GPR173) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.
This ELISA Test Kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Human GPR173 antibody. GPR173 present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Human GPR173 Antibody is added and binds to GPR173 in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated GPR173 antibody. After incubation unbound Streptavidin-HRP is washed away during a washing step. Substrate solution is then added and color develops in proportion to the amount of Human GPR173. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.
Serum Allow serum to clot for 10-20 minutes at room temperature. Centrifuge at 2000-3000 RPM for 20 minutes.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C within 30 minutes of collection.
Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for approximately 20 minutes. When collecting pleuroperitoneal fluid and cerebrospinal fluid, please follow the procedures above-mentioned.
Cell Culture Supernatant Collect by sterile tubes when examining secrete components. Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect the supernatants carefully. When examining the components within the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the cell concentration of approximately 1 million/ml. Damage cells through repeated freeze-thaw cycles to let out the inside components. Centrifuge at 2000-3000 RPM for approximately 20 minutes.
Tissue Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly and weigh before homogenization. Mince tissues and homogenize them in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20 minutes.
*Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample matrix interference may falsely depress the specificity and accuracy of the assay.
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour at 37°C.
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
Calculation of Result
Construct a standard curve by plotting the average OD for each standard on the vertical (Y) axis against the concentration on the horizontal (X) axis and draw a best fit curve through the points on the graph. These calculations can be best performed with computer-based curve-fitting software and the best fit line can be determined by regression analysis.
Contact Person: Lee