Beijing Exenatide (Exenatide) ELISA detection kit instructions, spot Exenatide elisa price Exenatide (Exenatide) ELISA kit instructions for use This kit is for research use only. 1 Purpose: This kit is used for the quantitative detection of Exenatide residues in related liquid samples such as serum, plasma and urine samples. Beijing Exenatide (Exenatide) ELISA detection kit instructions, spot exenatide elisa price experiment principle This kit uses a competitive ELISA method, coated with Exenatide (Exenatide) conjugated antigen in a microplate, added Exenatide standard or sample, free Exenatide and Exenatide conjugated antigen pre-coated on the microwell strip compete with each other for anti-Exenatide antibody Enzyme markers, developed with TMB substrate, the color changed from blue to yellow after adding the stop solution, detected with a microplate reader at a wavelength of 450nm, the absorbance value was inversely proportional to the content of tetracycline in the sample, and the standard curve was used to calculate the sample Exenatide (Exenatide) content. 3 Kit composition 3.1 Preencapped Exenatide (Exenatide) conjugated antigen removable enzyme labeling plate: 1 piece (12 wells × 8 strips). 3.2 Exenatide (Exenatide) standard products: 6 bottles (1ml / bottle), the contents are: 0pg / ml, 2 pg / ml, 6pg / ml, 18pg / ml, 54pg / ml, 162 pg / ml3.3 Anti-Exenatide antibody conjugate: 1 bottle (6ml). 3.4 Color developing solution A: 1 bottle (6ml). 3.5 Color developing solution B: 1 bottle (6ml). 3.6 Stop solution: 1 bottle (6ml), 2M sulfuric acid. 3.7 Sample diluent: 1 bottle (10 ×, 6ml), used for sample dilution. 3.8 Concentrated washing solution: 1 bottle (20 ×, 20ml) for washing plates. 3.9 A manual. 4 Materials needed but not provided 4.1 Equipment 4.1.1 Microplate reader with wavelength 450nm. 4.1.2 Crusher. 4.1.3 Measuring cylinder. 4.1.4 Oscillator. 4.1.5 Funnel. 4.1.6 Whatman No 1 or equivalent filter paper. 4.1.7 Micro pipette. 4.2 Reagents 4.2.1 Deionized water or distilled water. 4.2.2 Methanol. 5 Storage 5.1 The kit is stored at 2 ~ 8 ° C, do not freeze 5.2 Unused microplates should be sealed and stored dry 6 Cautions 6.1 Please read the instructions carefully before using the kit. 26.2 Do not use expired kits. 6.3 Before using the kit, return the reagent to room temperature (25 ± 2 ℃). It is recommended to return to temperature for at least 2 hours. 6.4 The standard product contains Exenatide (Exenatide), special care should be taken when using, and gloves should be worn during operation. 6.5 The stop solution contains sulfuric acid, which prevents burns to the skin and corrosion of clothing when used. 6.6 The tips used for different standards and samples can not be mixed, otherwise it will affect the test results. 6.7 The reagents in different batch kits should not be mixed; the tips used for different standards and samples should not be mixed, otherwise it will affect the experimental results. 6.8 The sample dilution in this kit must be used when diluting the sample, otherwise it will affect the experimental results. 6.9 Avoid blistering when mixing reagents. 7 Prepare 7.1 Exenatide standard working solution: 0pg / ml, 2 pg / ml, 6 pg / ml, 18pg / ml, 54pg / ml, 162 pg / ml 7.2 Concentrated washing solution: use distilled water Dilute at 1:20 (1 + 19) for standby 7.3 Sample diluent: reserved 7.3 Developer: reserved, avoid direct light 7.4 Reaction stop solution: reserved 8 Sample processing: (Sample should be strict during the extraction process Operate according to the instructions. The extraction process should be accurately diluted, otherwise the results will be inaccurate, and the sample should be stored in a cool and dark place and stored in cold storage) General sample processing 8.1 Take 10g of crushed sample, add 20ml 70% methanol solution 8.2 Strong oscillation 3 minutes 8.3 Filter with Whatman No 1 filter paper 8.4 Take 100 μl of the processed sample, add 400 μl of sample diluent 8.5 Take 100 μl of the diluent for analysis of animal tissue pretreatment 8.6 Accurately weigh 1 ± 0.05 g of homogenized tissue sample to 50 ml Into a polystyrene centrifuge tube; add 3ml of acetonitrile-acetone extraction solution, shake vigorously at 2000rpm for 20s, and then centrifuge at 4000r / min for more than 5min. , 750rpm vortex 20s8.9 take 100? L For the analysis of Beijing Exenatide (Exenatide) ELISA test kit instructions, spot exenatide elisa price feed pretreatment method 8.10 Accurately weigh 1 ± 0.05 g of crushed feed sample into a 50 ml polystyrene centrifuge tube; Add 4ml of acetonitrile, 1ml of acetone, shake vigorously at 2000rpm for 20s, and centrifuge at 4000r / min for 3min. 8.11 Take 0.7ml of supernatant and blow dry under nitrogen flow at 50 ° C. Add 2.8mL of sample diluent, vortex for 20s, mix for 100s ? l for the analysis of milk pretreatment method 8.13 take 1 ml of milk sample into a 5 ml polystyrene centrifuge tube; add 4ml of sample diluent, mix well and take 100? l for the analysis of milk powder pretreatment method 8.14 accurately weigh 0.3 g Milk powder sample into a 7 ml polystyrene centrifuge tube; add 2 ml of PBS solution, 2 ml of n-hexane, shake and mix for 8.154000r / min or more and centrifuge for 5 minutes to remove the organic layer and the middle layer, and remove the lower layer solution from 100 to 400 ? l sample diluent 8.16, mix and take 100? l for analysis 39 enzyme-free analysis steps 9.1 experimental instructions 9.1.1 before the experiment, please fully restore all reagents to room temperature (25 ± 2 ℃) outside the box, the time is about 2 hours. After warming to room temperature (25 ± 2 ℃), take out the microporous strips again. Re-seal the microporous strips and dry immediately at 2 ~ 8 ℃. Note: Make sure that the temperature is sufficient, otherwise the accuracy and accuracy of the test will be affected. 9.1.2 Please put the reagents back to 2 ~ 8 ℃ for storage immediately after use 9.1.3 Please do not change the analysis program 9.1.4 Please use accurate micro pipette 9.1.5 Once the operation starts, please do not interrupt any program 9.1.6 The reproducibility of ELISA results depends greatly on the operating procedures, please strictly follow the requirements 9.1.7 To avoid cross-contamination, each standard and sample should be loaded with a different tip 9.1.8 When loading Do not let the tip touch the solution or inner surface in the microwell. 9.2 Analysis step 9.2.1 Number in advance to mark the position of B0, standard and sample. It is recommended to perform double-well detection. 9.2.2 Take the required number of microwells (micro Hole strips are removable), reseal the excess strips and immediately put them back at 2 ~ 8 ℃ to store 9.2.3 Sample diluent and concentrated washing solution (20 ×) are diluted into working solution (diluted with distilled or deionized water) 9.2.4 Add 50 μl of 0.0 pg / ml standard solution to well B0. 9.2.5 Add 50 μl of standard solution to each standard well 9.2.6 Add 50 μl of sample solution to each sample well 9.2.7 Add 50 μl of all wells to all wells Anti-Exenatide antibody conjugate 9.2.8 Gently shaking reaction A few seconds. 9.3 Warm bath at 37 ° C for 30min (Tap the reaction plate from time to time during the warm bath to reduce the error of the two wells) 9.3.1 Shake off the liquid in the well and wash the microplate 5 times with the washing solution. The last time should be tapped on the absorbent paper to complete Remove the liquid from the hole. 9.4 Reaction 9.4.1 After the washing procedure is completed, immediately add 50 μl of chromogenic solution A and then 50 μl of chromogenic solution B to each microwell with a micropipette; shake the reaction plate slightly to mix thoroughly 9.4.2 37 ° C Warm bath 10min 9.4.3 Add 50μl of stop solution to each well, mix evenly 9.4.4 Detect the absorbance at 450nm, the result is read within 5min. 10 Result calculation 10.1 Quantitative analysis 10.1.1 The average value (B) of the absorbance value of each concentration standard solution and sample obtained is divided by the absorbance value (B0) of the first standard (0 standard) and multiplied by 100%, ie Percent absorbance value. B—the average absorbance value of the standard solution or the sample solution. The average absorbance value of the B0—0 ppb standard solution is 10.1.2. The logarithmic value of the Exenatide concentration is the X axis, and the percent absorbance value is the Y axis. Graph. According to the percent absorbance of the sample, the abscissa of the corresponding point can be obtained from the curve, which is the logarithm of the concentration of Exenatide, and the antilogarithm is the concentration of Exenatide in the measurement solution C (Ppb) 10.1.3 Because the sample has been pre-diluted, the sample concentration obtained from the standard curve must be multiplied by its dilution factor. 10.2 Semi-quantitative determination 10.1.1 Visual semi-quantitative determination: First select an appropriate standard solution to run with the sample, and determine whether the sample concentration value is less than or greater than the standard value based on the comparison of the absorbance value of the sample and the standard.
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