Manual of human NGALelias kit

Enzyme-linked immunoassay of human neutrophil gelatinase-associated lipocalin (NGAL)
Kit instruction manual
This kit is for research use only.
Detection range: 96T
0.2μg / L -8μg / L
purpose of usage:
This kit is used to determine the content of neutrophil gelatinase-associated lipocalin (NGAL) in human serum, plasma and related liquid samples. Experimental principle

This kit uses the double antibody sandwich method to determine the level of human neutrophil gelatinase-associated lipocalin (NGAL) in the specimen. Microporous plates were coated with purified human neutrophil gelatinase-associated lipocalin (NGAL) antibody to make solid-phase antibodies, and neutrophil gelatinase-related lipids were added to the microwells coated with monoclonal antibody in turn The carrier protein (NGAL) is combined with HRP-labeled neutrophil gelatinase-related lipocalin (NGAL) antibody to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The color depth is positively correlated with neutrophil gelatinase-associated lipocalin (NGAL) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human neutrophil gelatinase-related lipocalin (NGAL) in the sample was calculated by a standard curve.

Kit composition

1

30 times concentrated washing solution
20ml × 1 bottle
7
Stop solution
6ml × 1 bottle
2
Enzyme reagent
6ml × 1 bottle
8
Standard product (16μg / L)
0.5ml × 1 bottle
3
Enzyme coated plate
12 holes × 8
9
Standard dilution
1.5ml × 1 bottle
4
Sample diluent
6ml × 1 bottle
10
Instructions
1 serving
5
Developer A liquid
6ml × 1 bottle
11
Sealing film
2 sheets
6
Developer B liquid
6ml × 1 / bottle
12
sealed bag
1

Specimen requirements
1 The specimens are extracted as soon as possible after collection. The extraction is carried out according to relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided
2 NaN3 samples, because NaN3 inhibits horseradish peroxidase (HRP) activity.
Steps
1. Dilution of standard products: This kit provides one original standard product. The user can perform dilution in a small test tube according to the following chart.
2. Sample addition: set up blank wells (without adding samples and enzyme-labeled reagents in the blank control wells, the rest of the operations are the same), standard wells, and sample wells to be tested. Accurately add 50μl of the standard on the enzyme-coated plate, add 40μl of sample diluent to the sample well, and then add 10μl 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 ° C for 30 minutes.
4. Mixing solution: Dilute 30 times concentrated washing liquid with distilled water 30 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 50μl 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 50μl of developer A to each well, then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes.
10. Termination: Add 50μl 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 blank air conditioner zero and 450nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution.

8μg / L

Standard No. 5
150μl of original standard is added to 150μl standard dilution
4μg / L
Standard 4
150μl Standard No. 5 is added to 150μl standard dilution
2μg / L
Standard 3
150μl Standard No. 4 is added to 150μl Standard Diluent
1μg / L
Standard No. 2
150μl Standard No. 3 is added to 150μl Standard Diluent
0.5μg / L
Standard 1
150μl Standard No. 2 is added to 150μl Standard Diluent

Calculation

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; multiply by the dilution factor; or use the concentration of the standard 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. Precautions

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 sample 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 and then determine it. Multiple (× n × 5).

5. The sealing film is limited to one-time use to avoid cross-contamination.

6. Please keep the substrate away from light.

7. Strictly follow the instructions, and the test results must be determined by the microplate reader.

8. All samples, washing liquids and various wastes should be treated as infectious agents.

9. The components of different batches of this reagent shall not be mixed.

10. If there is any difference with the English manual, the English manual shall prevail.

Storage conditions and validity period

1. Kit storage :; 2-8 ℃.

2. Validity: 6 months

If you have any questions about the product, you can contact our company by phone, email, QQ, we ensure that you get a satisfactory response in the shortest time. We will continue to provide new products and new technology information, and will use the network platform to enable you to know our company, understand our products, and obtain technical support.

The appeal was the friendly shape of the blade: 5 to 7 inches long, with a rounded front edge and a boxier build than the typical chef`s knife, which usually stretches between 8 and 10 inches long and has a sleeker profile and a sword-like point. The style was developed for postwar Japanese home cooks as a more versatile alternative to vegetable cleavers- santoku reportedly means [three virtues," which are described variously as [meat, fish, and vegetables," or [chopping, slicing, and dicing"-and quickly became the country`s most popular Kitchen Knife.

Santoku Knife

Santoku Knife, 7-Inch Santoku Knife, Japanese Knife

Liveon Industrial Co.,Ltd. , https://www.liveonkitchen.com

This entry was posted in on