This reagent is intended for research use only and is designed to quantify vitellogenin (VTG) levels in shrimp serum, plasma, and related liquid samples. The assay is based on the double-antibody sandwich ELISA method. A solid-phase antibody, pre-coated onto a microwell plate, specifically binds to VTG present in the sample. Subsequently, an HRP-labeled VTG antibody is added, forming an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is introduced, which turns blue under the action of HRP and then changes to yellow when an acidic stop solution is added. The intensity of the color is directly proportional to the concentration of VTG in the sample. The absorbance is measured at 450 nm using a microplate reader, and the actual VTG concentration is determined by comparing the sample's OD value with a standard curve.
The kit includes a 48-well or 96-well configuration, along with essential reagents such as a coated enzyme-labeled plate, standard solutions, dilution buffers, substrates, and wash solutions. All components should be stored at 2–8°C. Proper storage ensures stability and performance over the shelf life of six months.
Sample preparation varies depending on the type of specimen. For serum and plasma, centrifugation is required after collection. Urine samples should also be centrifuged, while cell culture supernatants and tissue homogenates need additional processing steps, including dilution, homogenization, and centrifugation. It is crucial to avoid repeated freezing and thawing and to store samples at -20°C if testing is delayed. Samples containing sodium azide (NaN3) should not be used, as it may interfere with the HRP enzyme activity.
During the procedure, standards are prepared in serial dilutions, followed by sample addition, incubation, washing, and color development. The reaction is stopped, and absorbance is read within 15 minutes. Accurate pipetting, proper sealing, and careful handling are essential to ensure reliable results. A standard curve must be generated each time, and samples with high VTG concentrations should be diluted appropriately before testing.
The kit demonstrates excellent linearity, with a correlation coefficient (R) of at least 0.92. Intra- and inter-assay variability remain below 9% and 15%, respectively. The detection range spans from 3 ng/ml to 100 ng/ml. Always follow the manufacturer’s instructions closely, and treat all waste materials as biohazardous. Do not mix reagents from different batches. In case of discrepancies between the English and other language manuals, the English version takes precedence.
Easter Wooden Decorations Products
Ningbo Honor Imp & Exp Co.,Ltd , https://www.honormaket.com