Human soluble amyloid precursor protein β- w, ( hypersensitive ) detection kit
(Japan IBL imported, article number: 27732)
Japan's IBL China exclusive agent "Shenzhen Kerunda Biological Engineering Co., Ltd." dedicated to serve you
Introduction to the introduction
Alzheimer's disease (AD) was first published by German neuropathologist A. Alzheimer in 1907 and is recognized as a major cause of dementia.
principle
The solid phase ELISA kit uses two highly specific antibodies. The TMB substrate solution is added as a color developer. The intensity of color development is related to the amount of human soluble amyloid precursor protein β-w (sAPP β-w). In proportion to.
examination range
0.78 - 50 ng/mL
expected usage
For research purposes, not for diagnostic procedures.
■This IBL test kit can be used for quantitative detection of sAPPβ-w in human serum, EDTA plasma, cerebrospinal fluid, and cell culture supernatant.
â– It is recommended that EDTA plasma samples be diluted more than 4 times.
â– It is recommended to dilute cerebrospinal fluid more than 8 times
â– The supernatant of the cell culture medium, such as FCS, may cross-react. Therefore, it is recommended to set the negative property control.
Kit composition
1 pre-coated plate: anti-human sAPPβ-w rabbit IgG monoclonal antibody, affinity purification 96T
2 concentrated enzyme-labeled antibody (enzyme conjugate): 30 × 0.4 mL x 1
3 standard: recombinant human sAPPβ-w protein 0.5mL x 2
3 standard: recombinant human sAPPβ-w protein 0.5mL x 2
5 enzyme conjugate dilution 12mL x 1
8 concentrated washing solution: 40 × 50mL x 1
Instructions
1 required experimental equipment ( but not provided in this kit )
Microplate reader (450nm) micropipette and nozzle
Measuring cylinder and beaker
Refrigerator (4 ° C) coordinate paper (log / log)
Absorbent paper
Washing bottle
Disposable test tube for "2 concentrated enzyme-labeled antibody" and "6 substrate solution: TMB solution"
2 preparation
1) Preparation of washing liquid
"8 concentrated washing liquid" is a 40 times concentrated liquid. After being placed at room temperature, 50 mL of the concentrated solution is dissolved in 1950 mL of deionized water (ie diluted 1:40) to prepare a diluted washing buffer. Can be stored in the refrigerator for 2 weeks.
2) Preparation of enzyme conjugates
"2 concentrated enzyme-labeled antibody " is a 30-fold concentrated enzyme conjugate, which is diluted with a "5 enzyme conjugate" using a single test tube to prepare the desired enzyme conjugate solution. The required amount is diluted 1:30)
3) Preparation of standard products
Add 0.5 mL of deionized water to the bottled “3 standard †and mix to make 100 ng/mL human sAPPβ-w standard.
4) Dilution of standards
Prepare 8 tubes for standard dilution and number. Add 230 μL of “4EIA buffer†to each tube. Take 230 μL of standard solution in tube 1 and mix. Then take 230 μL from tube 1. Put it into the test tube 2, according to this rule, as shown in the following figure, prepare a series of standard products. The 8th test tube is 0 ng/mL as a zero standard.
5) Dilution of the sample
The sample must be diluted with the “4EIA buffer†provided in the kit . If the concentration range of sAPPβ-w in the sample is not clear, it is recommended to pre-form several different concentrations to determine the appropriate concentration.
Detection step
All reagents are equilibrated to the greenhouse (approximately 30 minutes) and mixed before use. Ensure that the reagents have no quality problems. Prepare the standard and prepare a standard curve while testing the sample.
The test flow chart is as follows:
1 Set the reagent blank control. Add 100 μL of “4EIA buffer†to the microwell.
2 Add 100 μL sample blank (tube 8), series standard (tube 1-7), and sample in the corresponding holes.
3 Incubate overnight at 4 ° C after the cover
4 Wash the plate with a washing bottle. Then add the washing buffer, no need to cover for 15-30 seconds, then completely remove the washing buffer. This step needs to be repeated at least 7 times. Finally, remove all residues on the absorbent paper. Droplet.
If the automatic washing machine is used, after the plate is washed 4 times on the washing machine, the above manual washing step still needs to be repeated 3 times.
5 add 100 μL of enzyme solution in each well.
6 cover and incubate at 4 ° C for 30 minutes
7 Wash the plate 9 times in step 4.
8 Add the required amount of “6 substrate solution: TMB solution†to a disposable tube. Then take 100 μL of substrate solution in each well. Note that the excess substrate in the disposable tube should not be returned. Avoid cross infection in the bottle.
9 Incubate the coated plate for 30 minutes at room temperature. After adding the substrate solution, the liquid in the well will turn blue.
10 Add 100 μL of “7 Stop Liquid†to each well, and mix the light-strip strips. After adding the stop solution, the liquid in the well will turn yellow.
11 Remove the dirt or droplets at the bottom of the plate to ensure no bubble formation. Measure the absorbance at 450 nm with a microplate reader within 30 minutes after the addition of the stop solution.
pay attention
1 The sample should be tested immediately after collection. If it is stored frozen, it needs to be thawed to avoid repeated freezing and thawing.
2 samples must be diluted with "4EIA buffer".
3 recommended sample and each standard for double detection
4 samples should be kept in the neutral pH range. Because the contamination of organic solvents will affect the test results
5 wash buffers provided with the kit can be used for washing plates. Insufficient washing may result in erroneous results.
6 Light the plate on the absorbent paper to remove residual droplets. Do not scratch the micropores with absorbent paper.
7 "6 substrate liquid" should be stored in the dark to avoid contact with metal.
8 After adding “7 Stop Liquidâ€, it is necessary to read within 30 minutes.
Result analysis and calculation
All measured absorbance values ​​(including standards, unknown test samples) are subtracted from the absorbance values ​​of the sample blanks. On the coordinate paper, the standard concentration is used as the abscissa and the corresponding absorbance is taken as the ordinate. The point is drawn on a double logarithmic scale paper. The concentration of the test sample can be read directly on the standard curve.
The above standard curve is for demonstration purposes only and cannot be used for reading sample results. It is necessary to establish a standard curve for each test.
This translation is for reference only, please refer to the original for details.
Exclusive distributor in China: Shenzhen Kerunda Bioengineering Co., Ltd.
Company address : 6th Floor, No. 10, Yanshan Road, Shekou, Nanshan District, Shenzhen. Zip code : 518067
Contact number (8 lines) Fax : +86 755 26814431
Free ordering phone .
Technical consultation : landline, 0755-26680196; mobile phone, 15711973608; E-mail,.
Official website :
Welcome to request detailed information, online email:
Electronic cigarette is electronic product that generates vapor feel, taste like as a cigarette. Nic does not cause cancer, what really causes cancer are the thousands of harmful chemicals generated during tobacco burning. E-cigarette work as harming reducing product, the vapor contains fewer toxic chemicals in lower concentrations than traditional tobacco cigarette.
E-cigs also have name vape pen, disposable cigarette, it has many flavors such as apple, peach, orange, grape etc, people can enjoy multi-flavored Nic delivery experience through vapor, instead of Tobacco.
Axiswell is a manufacturer of no leak electronic cigarette vape with R&D, production and sales team. Our star products are disposable vape pen, closed pod system, vape pods. Our MOQ is low and we provide OEM/ODM service. We understand good taste and reasonable pricing are important for long term cooperation between us and clients. We wish to cooperate with every clients no matter big quantity or small quantity, reaching win-win outcome!
Electronic cigarette,vapes e cigarette,disposable electronic cigarette,vape electronic cigarettes,electronic cigarettes vape
Shenzhen Axiswell Technology Co., Ltd , https://www.medhealthycare.com