Rat insulin receptor (Insulin R) ELISA kit instruction manual

Shanghai Xitang Biotechnology Co., Ltd. 021-55229872, 65333639
Rat insulin receptor (Insulin R) ELISA kit instruction manual
  ( used in serum, plasma, cell culture supernatants and other biological fluids )
principle
This experiment used double antibody sandwich ABC-ELISA. The anti-rat Insulin R monoclonal antibody was coated on the microtiter plate, the standard and the sample of Insulin R were combined with the monoclonal antibody, and biotinylated anti-rat Insulin R was added to form an immune complex attached to the plate. The root peroxidase-labeled Streptavidin is combined with biotin, the substrate working solution is blue, and finally the stop solution sulfuric acid is added. The OD value is measured at 450 nm. The Insulin R concentration is proportional to the OD value, which can be obtained by drawing a standard curve. Insulin R concentration in the specimen.
Kit composition ( 2-8 ° C preservation)
Coated Wells
96 holes
Enzyme Conjugate
12ml
10× specimen dilution (Sample Buffer)
12ml
20×Wash Buffer
50ml
Standards: 50ng/bottle
2 bottles
Substrate working fluid (TMB Solution)
12ml
Primary antibody working solution (Biotinylated Antibody)
12ml
Stop Solution
12ml
Prepare reagents and collect blood samples
1. Collect specimens: serum, plasma (EDTA), cell culture supernatant, tissue homogenate, etc., as soon as possible, store at 2-8 ° C for 48 hours; longer time must be frozen (-20 ° C or -70 ° C) to avoid Repeated freezing and thawing. 1 with sample dilution is normal before the sample measurement: 10 dilution (take 20ul, sample plus diluent 180ul, diluted 10 times).
2. Standard solution preparation: Add 0.5 ml of distilled water before use and mix to form a 100 ng/ml solution. Set the standard tube 8 tube, the first tube plus the standard dilution 900ul, the second to the eighth tube to add the sample dilution 500ul. Add 100 ul of 100 ng/ml standard solution to the first tube, mix and aspirate 500 ul with the sampler, and transfer to the second tube. Repeat the dilution as described above, and remove 500 ul from the seventh tube and discard it. The eighth tube is a blank control.
3. The 10× specimen dilution was diluted 1:10 with distilled water (example: 1 ml concentrated dilution + 9 ml distilled water).
4. Washing solution: diluted 1:20 with distilled water (example: 1 ml concentrated washing solution added to 19 ml of distilled water)
Test procedure
1. Loading: Add 100 ul of standard or sample to be tested in each well. Mix the reaction plate thoroughly and let it stand at 37 °C for 120 minutes.
2. Wash the plate: Wash the plate thoroughly with washing solution 4-6 times, and dry it on the filter paper.
3. Add 100 ul of the first antibody working solution to each well. The reaction plate was thoroughly mixed and placed at 37 ° C for 60 minutes.
4. Wash the board: the same as before.
5. Add 100 ul of enzyme-labeled antibody working solution per well. The reaction plate was placed at 37 ° C for 30 minutes.
6. Wash the board: same as before.
7. Add 100 ul of substrate working solution to each well and let it react at 37 ° C for 15 minutes in the dark.
8. Add 100 ul of stop solution to each well and mix.
9. Measure the absorbance at 450 nm using a microplate reader within 30 minutes.
Result calculation and judgment
1. All OD values ​​should be subtracted from the blank value before calculation.
2. Using standard products 10, 5, 2.5, 1.25, 0.625, 0.312, 0.156, 0 ng/ml as the abscissa and OD as the ordinate, plot on the coordinate paper and draw the standard curve.
3. Find the corresponding Insulin R content on the graph based on the OD value of the sample, and multiply by the dilution factor.
Kit performance
1. Sensitivity: The minimum Insulin R detection concentration is less than 0.1 ng/ml.
2. Specificity: Recombinant or natural rat Insulin R can be detected simultaneously. Does not cross-react with other cytokines in rats.
3. Repeatability: The coefficient of variation in both the plate and the plate is less than 10%.
Precautions
1. It is recommended to make double holes for the above standard holes and samples to be tested. The standard curve should be made at the same time for each measurement.
2. The washing process is critical. Insufficient washing will result in an accuracy error and an erroneous rise in the OD value.
3. After the slats are opened, the remaining slats should be sealed again to keep the slats dry .
4. This kit should be stored in a 4oC refrigerator.
5. This kit is for scientific research only and cannot be used for clinical diagnosis!

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Self-cleaning filter is a kind of filter that directly intercepts impurities in water, removes suspended matter, particulates, reduces turbidity, purifies water quality, reduces system fouling, bacteria and algae, rust and so on. In order to purify the water quality and protect the system other equipment work normally, the water enters the self-cleaning filter body from the intake, because of the intelligent (plc,pac) design, the system can automatically identify the impurity deposition degree, Automatic full discharge for discharge valve signal. Sinofiltec all series filtration solution can meet kinds of different water quality in industry area, our professional system design is made and provided according to each customer`s requirement to solve water treatment efficiently, by global sales network and close cooperation with customers, our manufacture capability, rich experience and technology ability can help you reduce costs, reduce energy consumption, improve the quality.

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The water enters the filter through the inlet, and the impurity of the larger particle is first filtered by the coarse filter element assembly, then the impurity of the fine particle is filtered through the fine filter net, and the water is discharged from the outlet after the impurity of the fine particle is filtered by the fine filter net. In the filtration process, the inner layer of the fine filter gradually accumulates, and a pressure difference between the inner and outer sides of the filter is formed. When this pressure difference reaches the preset value, the automatic cleaning process begins: the drain valve is opened, the hydraulic motor chamber and the hydraulic cylinder in charge of the assembly release pressure and drain the water; The pressure in the hydraulic motor chamber and the suction pipe is greatly reduced.

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