Shanghai Lianshuo Biological Technology Co., Ltd. is one of the main suppliers of immunology products in China. It provides various original and sub-packaged ELISA kits with high quality and low price. You provide technical service and support, and any problems with the kit can help you solve it, eliminating your worries. And it can be used for free testing, so that your experiment saves time and effort, and better serves you. If you have any other questions, please contact us in time.
Instruction Manual for Rat S-100B Protease Linked Immunoassay (ELISA) Kit
Specifications: original / 96T 48T / 96T
This kit is used to determine the content of S-100B protein in rat serum, plasma and related liquid samples.
Specimen processing and requirements
1. Serum and plasma samples can be directly measured;
2. Urine, cerebrospinal fluid, and intraperitoneal fluid: After 2000-3000 rpm / separation of the heart for 10 minutes, take the supernatant for testing.
3. The samples containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity.
4. Experiment as soon as possible after collection. If the experiment cannot be performed immediately, the specimen can be stored at -20 Â° C, but repeated freezing and thawing should be avoided.
20 times concentrated washing liquid
30ml Ã— 1 bottle
6ml Ã— 1 bottle
6ml Ã— 1 bottle
Standard product (180ng / L)
0.5ml Ã— 1 bottle
Enzyme coated plate
12 holes Ã— 8
1.5ml Ã— 1 bottle
6ml Ã— 1 bottle
Developer A liquid
6ml Ã— 1 bottle
Developer B liquid
6ml Ã— 1 / bottle
Bring your own items
1. 37 â„ƒ thermostat
2. Standard specification microplate reader
3. Precision pipettes and disposable tips
4. Distilled water
5. Disposable test tubes
6. Absorbent paper
1. Dilution and loading of standard products: 10 standard wells are set on the enzyme-coated plate, 100Î¼l of the standard products are added to the first and second wells respectively, and then the standard products are added to the first and second wells 50Î¼l of diluent, mix well; then take 100Î¼l from the first well and the second well and add them to the third and fourth wells respectively, and then add 50Î¼l of standard diluent to the third and fourth wells respectively, mix well; Then take 50Î¼l each in the third and fourth wells and discard it, then add 50Î¼l each to the fifth and sixth wells, and then add 50ul of the standard dilution solution to the fifth and sixth wells respectively, and mix well; After mixing, take 50Î¼l from the fifth and sixth wells and add them to the seventh and eighth wells respectively. Then add 50Î¼l of the standard dilution solution to the seventh and eighth wells respectively. Take 50Î¼l from the eight wells and add them to the ninth and tenth wells. Then add 50Î¼l of the standard dilution solution to the ninth and tenth wells. After mixing, take 50Î¼l from the ninth and tenth wells and discard. (After dilution, the volume of each well is 50Î¼l, and the concentrations are 120ng / L, 80ng / L, 0ng / L, 20ng / L, 10ng / L).
2. Adding samples: set up blank wells (the blank control wells do not add samples and enzyme reagents, the rest of the steps are the same) and the sample wells to be tested. Add 40Î¼l of sample diluent to the test sample well of the enzyme-coated plate, and then add 10Î¼l of the sample to be tested (the final dilution of the sample is 5 times). Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix.
3. Incubation: seal the plate with a sealing film and incubate at 37 Â° C for 30 minutes.
4. Mixing solution: dilute 20 times concentrated washing liquid with distilled water 20 times and reserve
5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing solution, let it stand for 30 seconds and then discard, repeat this 5 times, pat dry.
6. Add enzyme: add 50Î¼l of enzyme label reagent to each well, except blank well.
7. Incubation: The operation is the same as 3.
8. Washing: The operation is the same as 5.
9. Color development: Add 50Î¼l of developer A to each well, and then add 50Î¼l of developer B, mix gently, and develop at 37 Â° C in the dark for 15 minutes.
10. Termination: Add 50Î¼l of stop solution to each well to stop the reaction (at this time, the blue color turns to yellow).
11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution.
Taking the concentration of the standard as the abscissa and the OD value as the ordinate, draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; then multiply by the dilution factor; Calculate the linear regression equation of the standard curve with the OD value, substitute the OD value of the sample into the equation, calculate the sample concentration, and multiply it by the dilution factor to obtain the actual concentration of the sample.
1. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. If the enzyme label coated plate is unopened, the strip should be stored in a sealed bag.
2. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in a water bath during dilution, and the results will not be affected during washing.
3. The sampler should be used at each step of sample addition, and the accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples.
4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the test substance in the specimen is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute it with a certain multiple (n times) of the sample diluent before measuring, and finally multiply the total dilution when calculating Multiple (Ã— n Ã— 5).
5. Strictly follow the instructions, and the test results must be determined by the reading of the microplate reader.
6. The components of different batches of this reagent shall not be mixed.
7. The sealing film is limited to one-time use to avoid cross-contamination.
8. All samples, washing liquids and various wastes should be treated as infectious agents.
9. Please keep the substrate away from light.
8ng / L-150ng / L
Storage conditions and validity period
1. Kit storage :; 2-8 â„ƒ.
2. Validity: 6 months
Spoons are one of the essentials in every kitchen, no one can set a meal ready without using it. We here display many other functional spoons except traditional spoons. Except for daily meal cooking, they are adopted in the condition of coffee serve or honey stirring. Ice cream scoops can be a good helper for heat releases in a hot summer afternoon, you may feel like getting out of the midsummer when a full scoop of ice cream appears in front of you.
Ice Cream Scoop
We have various functional designs to define different ice cream scoops. This widget, except the tranditional usage to make ice cream, has expanding functions in our daily life:
1. ingenious meal creator. You may put your child to feed himself/herself independently if you can create a cartoon shape meal, such as Mickey Mouse head.
2. fruit server. Use it as a server to pick out fruit balls, sush as watermelon and cantaloupe.
Are you still despressed when you are facing to decide how many condiment is needed? Take a look at the Measuring Tools, they are useful in both daily cooking and bakery. It's also a good helper for dieters that can prevent any food waste.
An essential in morden kitchen life. Different dessert spoons for cakes, coffee and honey to choose from.
Cute Measuring Spoons,Liquid Measuring Cup,Dessert Spoon,Honey Server Spoon
V-Boom's Industrial Co.Ltd , https://www.v-booms.com