Eva Nail File,Mini Nail Files,Cute Nail Files,Professional Nail Files Bonet Houseware Co., Ltd. , https://www.manicureset.de
Sudan Red ELISA Kit Instructions for Use This kit is for research use only. 1 Use purpose Use purpose Use purpose Use purpose: :: This kit is used for quantitative detection of Sudan red residue in eggs, chili powder, and chili sauce. 2 Experimental principle Experimental principle Experimental principle Experimental principle This kit uses a competitive ELISA method, coated with Sudan red conjugated antigen in a microwell plate, added Sudan red standard or sample, free Sudan red and pre-coated on the microwell strip Sudan red conjugated antigens compete with each other for anti-Sudan red antibody enzyme markers, developed with TMB substrate, the color changes from blue to yellow after the addition of stop solution, and detected by a microplate reader at 450nm wavelength. The content of Sudan red is inversely proportional, and the content of Sudan red in the sample is calculated by the standard curve. 3 Kit composition Kit composition Kit composition Kit composition 3.1 Pre-coated Sudan Red conjugated antigen detachable enzyme label plate: 1 piece (12 wells × 8 strips). 3.2 Sudan Red Standard: 6 bottles (1ml / bottle), the contents are: 0 ppb, 0.1 ppb, 0.3 ppb, 0.9 ppb, 2.7 ppb, 8.1 ppb. 3.3 Anti-Sudan red antibody enzyme 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), used for washing plates. 3.9 A manual. 4 Needed but needed and needed but not provided Materials not provided Materials not provided Materials not provided Materials 4.1 Equipment 4.1.1 Wavelength 450nm microplate reader. 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 Storage Storage Storage 5.1 The kit is stored at 2 ~ 8 ℃, do not freeze 5.2 Unused microplates should be sealed and stored dry 6 Cautions Cautions Cautions 6.1 Please read the instructions carefully before using the kit. 6.2 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. 2 6.4 The standard contains Sudan Red. Special care should be taken when using it. 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 Working fluid preparation Working fluid preparation Working fluid preparation Working fluid preparation 7.1 Sudan red standard solution: 0ppb, 0.1ppb, 0.3 ppb, 0.9ppb, 2.7 ppb, 8.1ppb 7.2 Concentrated washing solution: 1:20 (1+ with distilled water) 19) Dilute for use 7.3 Sample diluent: Dilute with distilled water at 1:10 (1 + 9) for use 7.3 Developer: Used, avoid direct light 7.4 Reaction Stop Solution: Used 8 Sample handling procedures Sample handling procedures Sample handling Procedure Sample processing procedures (samples should be strictly operated in accordance with the instructions during the extraction process, and they should be accurately diluted during the extraction process, otherwise the results will be inaccurate, and the samples should be stored in a cool and dark place and refrigerated storage) ， Add 20ml 70% methanol solution 8.2 Vibrate vigorously for 3 minutes 8.3 Filter with Whatman No 1 filter paper 8.4 Take 25μl of the treated sample, add 25μl of sample dilution to the reaction well (sample dilution factor is 2) 9 Enzyme-free analysis step Analysis steps Enzyme-free analysis steps Enzyme-free analysis steps 9.1 Notes to the experiment 9.1.1 Before the experiment, please restore all the reagents to room temperature (25 ± 2 ℃) outside the box for 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), re-seal the excess strips and immediately put them back at 2 ~ 8 ℃ to store 9.2.3 Sample dilution (10 ×) 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 ng / ml standard solution to well B0 9.2.5 Add 50, l of standard solution to each standard well 9.2.6 Add 50 μl of sample solution 9.2.7 to each sample well Add 50 μl of anti-Sudan red antibody enzyme conjugate to all wells 9.2.8 Gently shake the reaction plate Seconds. 9.3 Warm bath at 37 ° C for 30min (tap the reaction plate from time to time during the warm bath to reduce double-hole errors) 9.3.1 Shake off the liquid in the well and wash the microplate 5 times with washing liquid. The last time should be tapped on absorbent paper Remove the liquid from the hole. 9.4 Reaction 3 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 Warm bath at ℃ for 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 result Calculation result calculation 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 then Multiply by 100% to get the percent absorbance value. B—The average absorbance value of the standard solution or the sample solution. The average absorbance value of the B0—0 μg / L standard solution is 10.1.2. Take the logarithmic value of the Sudan red concentration as the X axis and the percent absorbance value as the Y axis. According to the percent absorbance value of the sample, the abscissa of the corresponding point can be obtained from the curve, which is the logarithmic value of the concentration of Sudan red, and the antilogarithm is the concentration of Sudan red C (ppb) in the measurement solution. 10.1.3 Because the sample has passed Pre-dilute, so 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. 10.1.2 Instrument semi-quantitative determination: first select an appropriate standard solution to run with the sample, and determine whether the concentration value of the sample is less than or greater than the standard value according to the color depth comparison of the sample and the standard product. 11 Specificity Specificity Specific substance cross reaction Sudan red 100% 12 Kit parameters Kit kit parameters Kit parameters Kit kit parameters The lower detection limit of this kit is 0.05ppb B0 The optimal value of absorbance should be greater than 1.0 in the kit absorbance plate The error is less than 8%, and the error between boards is less than 15%. The recovery rate of the tissue sample extraction method provided in this specification is greater than 80%. 13 The standard curve range provided by the kit is 0.1ppb ~ 8.1ppb. 14 Analysis Limits Analysis Limitations Analysis Limitations Analysis Limitations The samples tested positive by this kit should be confirmed by another method such as HPLC or GC / MS.