Human endothelial nitric oxide synthase (eNOS) ELISA kit instruction manual

Posted on

Human endothelial nitric oxide synthase (eNOS) ELISA kit
  ( used in serum, plasma, cell culture supernatants and other biological fluids )
principle
This experiment used double antibody sandwich ABC-ELISA. The anti-human eNOS monoclonal antibody was coated on the microtiter plate, the eNOS of the standard and the sample were combined with the monoclonal antibody, and biotinylated anti-human eNOS was added to form an immune complex attached to the plate, horseradish peroxidase. The labeled Streptavidin is combined with biotin, and the substrate working solution is blue. Finally, the stop solution sulfuric acid is added, and the OD value is measured at 450 nm. The eNOS concentration is directly proportional to the OD value, and the eNOS concentration in the sample can be obtained by drawing a standard curve.
Kit composition ( 2-8 ° C preservation)
Coated Wells
96 holes
Enzyme Conjugate
12ml
10× specimen dilution (Sample Buffer)
12ml
20×Wash Buffer
50ml
Standards: 20ng/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) Save to avoid repeated freezing and thawing.
2. Standard solution preparation: Add 0.5 ml of distilled water before use and mix to form a 40 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 the standard solution of 40 ng/ml 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 per well, set 37 The reaction was carried out in the dark at °C for 15 minutes.
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. Take the standard products 4000, 2000, 1000, 500, 250, 125, 62.5, 0 pg/ml as the abscissa and OD as the ordinate. Draw on the coordinate paper and draw the standard curve.
3. Find the corresponding eNOS content on the graph based on the OD value of the sample.
Kit performance
1. Sensitivity: The minimum eNOS detection concentration is less than 30pg/ml.
2. Specificity: Recombinant or natural human eNOS can be detected simultaneously. Does not cross-react with other human cytokines.
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 4 o C refrigerator.
5. This kit is for scientific research only and cannot be used for clinical diagnosis!

Orange Enzyme Hand Soap

Orange soap with a clean, dried on the skin of course, is to remove the dirt dirt, skin secretions, excretions, chemicals or bacteria, etc. In today's emphasis on cosmetic results, remove the skin surface of cosmetics, skin care products or drugs is also a major one uses. Because in Orange soap he added perfume lotus, so have a navel orange aroma and also with the aroma of lotus.


Companies registered capital of 35 million yuan, the end of 2014 the total assets of 48.69 million yuan, including fixed assets of 37.52 million yuan. The company's existing cooperation Orange cultivation base 7043.5 acres, the company production base is located in Jiangxi County Tech Industrial Park Chu Tan industrial area, covers an area of 120 acres, it has built a standard plant 9,000 square meters, Nissan 6000 kg Orange enzymes and other liquid enzyme products. Enzyme, known as enzyme, refers to a polymer substance having biocatalytic functionality. In the catalytic reaction system an enzyme, the reactant molecules are known as substrates, enzyme substrates by catalytic conversion to another molecule. Almost all cellular activity of enzymes involved in the process are required to improve efficiency. Similar to other non-biological catalysts, enzymes chemical reactions by lowering the activation energy to accelerate the rate of the reaction, most of the enzyme catalyzed reaction rate can be increased a million times; in fact, the enzyme is to provide an activation energy needs than another low way, so that more particles to have less than the activation energy of the reaction kinetic energy, thus speeding up the reaction rate. Enzyme as a catalyst, in itself is not consumed during the reaction, it does not affect the chemical equilibrium reactions. Positive enzyme catalysis, but also a negative catalytic effect, not only to accelerate the reaction rate, but also to reduce the reaction rate. And other non-living catalysts is different, having a high degree of specificity of enzyme, only a catalytic reaction or produce a particular specific configuration.

Orange Enzyme Hand Soap,Pure Artificial Production Hand Soap,Orange Lotus Aromas Hand Soap,Cosmetic Whitening Skin Hand Soap

Ganzhou Green days Biochemical Technology Co., Ltd. , http://www.cn-gangdao.com