| Description | Product usage adviceIncubation temperature: 20-37 °C / 68-100 °FResponse time: Moderate (up to 30 minutes) at 20-37°CShelf life: 24 monthsActive ingredients and principle of actionIn different buffer solutions (pH = 9.5), different additives were added as required, and the active Product usage adviceIncubation temperature: 20-37 °C / 68-100 °FResponse time: Moderate (up to 30 minutes) at 20-37°CShelf life: 24 monthsActive ingredients and principle of actionIn different buffer solutions (pH = 9.5), different additives were added as required, and the active ingredient p-nitrophenyl phosphate (pNPP) was dissolved.Alkaline phosphatase transfers phosphate residues to acceptors. Under alkaline conditions, a yellow color will appear due to the formation of nitrophenols.Main advantageHigh absorbance yield comparable to freshly dissolved pNPP tabletsWide measuring rangelow background signalLow blank drift during long-term storage (<0.18 AU over 24 months)High color stability after quenching the reaction with used stop solutionStorage, Shipping and Filling InformationStore at 2-8°C protected from light. Under these conditions, the expiration date printed on the label is valid for unopened bottles.Spontaneous decay increases the background. The rate of decay increases if stored at room temperature. Therefore, storage and transport at room temperature should be avoided. However, the activity of the solution was not affected by storage at room temperature. The solution remains valid beyond the due date, but some applications, especially those that include visual assessments, may be hindered by increased background.Contaminated or leaked substrate solutions from damaged bottles should not be reused and must be destroyed.Ship in an isolated container with some refrigerated bags.The following bottling instructions should be considered:Work in a dust-free and dark room.Keep the solution as cool as possible.Avoid contact of solution with any metal parts.Thoroughly clean all instruments and containers.Wear powder-free gloves when bottling.Close the bottle immediately to minimize the effects of light and dust.Use an opaque, clean bottle made of HDPE or PP.General instructions for use in ELISAOnly qualified laboratory staff familiar with the basics of immunological methods should use these solutions.Substrate solutions can be used in qualitative and quantitative ELISA procedures.When using a 96-well microtiter plate, it is recommended to add 100 µL of pure substrate per well after incubation and washing. After substrate incubation, the reaction can be stopped and photometric measurements can be performed. Using a higher incubation temperature (37°C) may shorten the incubation time.stop 2. Further increases in signal cannot be safely ruled out using other commercially available stopping solutions. The addition of stop solution did not change the general shape of the spectrum.Unstopped and stopped solutions should be measured at 405 nm, background corrected: should be measured at 620 nm.WasteDisposal of waste must comply with national and local laws and regulations. Disposal of packaging must comply with national and local disposal regulations... Read More | Inquire | Inquire | ProductsThis product is a high purity genomic DNA extract from 293T cells, agarose gel (0.7%) electrophoresis showed that the size of the DNA extract is more than 15Kb, and basically no degradation, the product is ultimately preserved in TE Buffer, which can be widely used in molecular biology ProductsThis product is a high purity genomic DNA extract from 293T cells, agarose gel (0.7%) electrophoresis showed that the size of the DNA extract is more than 15Kb, and basically no degradation, the product is ultimately preserved in TE Buffer, which can be widely used in molecular biology experiments, such as PCR, enzyme digestion, hybridization, microarray analysis, and other molecular biology experiments.The product was quantified using NanoDrop One at a concentration of 200 ng/µL.Preparation and precautions before useLong-term storage at -20˚C is recommended. Before use, the bottle should be removed from the refrigerator and equilibrated to room temperature and centrifuged before opening the cap for use. Samples should be restored to the sealed state as soon as possible after opening.How to use (take qPCR experiment as an example)1. Amplification template preparationThe samples to be detected were diluted with TE (10 mM Tris-Cl, pH 8.0,1 mM EDTA), and the concentration after dilution was as close as possible to the range of 0.05-10 ng/µL. The samples were placed on ice at 4°C and set aside.2. Standard dilution: according to the following table, firstly dilute Human DNA Standard 1 (100ng/uL) with TE to make 5 different concentrations of standards according to the table below. 10ng/µL of DNA Standard 1 (Std. 1) can be stored stably at -20℃ for 1 month; Std2-5 can only be used on the same day, and should be placed at 4℃ or on ice when not in use for the time being after preparation. When not used temporarily after preparation, it should be stored at 4℃ or on ice.styleCorresponding concentration (ng/µL)Minimum dilution volume (in µL)Std.11010 [100 ng/µL DNA Standard 1] + 90 TEStd.22.520 [Std. 1] +60 TEStd.30.62520 [Std. 2] +60 TEStd.40.1562520 [Std. 3] +60 TEStd.50.039062520 [Std. 4] +60 TE3. qPCR reaction system preparationThe cryopreserved reagents to be used were completely thawed and mixed by inversion several times before preparation, and then briefly centrifuged and prepared for use. 20 µL of the base reaction system was as follows.The base reaction system for 20 µL was as follows:reagents20µL reaction system2×qPCRMix10µLPrimerMixXµLProbeMixXµLTemplate4µLddH2OMake up to 20 µLNote: High Rox model: add 1 µL of 50×High Rox per 50 µL of reaction system; Low Rox model: add 1 µL of 50×High Rox per 500 µL of reaction system.Usually, better results can be obtained with a primer concentration of 0.2 µM, and 0.1-1.0 µM can be used as a reference for setting the range.The concentration of the probe used is related to the fluorescent quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the manual of the instrument or the specific requirements for the use of each fluorescent probe for the adjustment of the concentration during actual use.Prepare a sufficient amount of reaction system mixture as required. After the reaction system has been prepared and mixed thoroughly, add 16 µL per well to the reaction wells. Then add the prepared standard and diluted sample into the corresponding reaction wells, the volume of addition is 4µL/well. TE was added to the blank control tube, and the same amount of TE was added at 4 µL/well.It is recommended to use 20 µL for the reaction, if you need to perform a smaller system reaction, reduce the system components in equal proportion.4. qPCR reaction programThe following is an example of our GoldStar Probe Mixture reaction conditions, which should be improved and optimized according to the PCR product template, primer structure and target fragment size.movetemptimingcirculatepremutability95°C10min1denaturation95°C10sec55Annealing/Extension60°C30sec5Data analysis1. Standard curve productionThe standard curve was plotted with reference to the Excel sheet for data processing. The correlation coefficient R2 of the standard curve should not be lower than 0.98, and the slope should be between -3.1 and -3.6 when the Ct value is the vertical coordinate. If the parameters of the standard curve are unreasonable, it is recommended to repeat the experiment... Read More | Purity>95% (SDS-PAGE) Endotoxin level<1.0 EU/µgFunctionInhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells |