Human CC Chemokine Receptor-1 (CCR-1) ELISA Kit Instructions for Use

Human CC Chemokine Receptor -1 (CCR-1) ELISA Kit

Â ( used in serum, plasma, cell culture supernatants and other biological fluids )

principle

This experiment used double antibody sandwich ABC-ELISA. The anti-human CCR-1 monoclonal antibody was coated on the microtiter plate, the CCR-1 in the standard and the sample was combined with the monoclonal antibody, and biotinylated anti-human CCR-1 was added to form an immune complex attached to the plate. Horseradish peroxidase-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 CCR-1 concentration is directly proportional to the OD value, and the standard can be drawn. The curve finds the concentration of CCR-1 in the specimen.

Kit composition ( 2-8 Â° C preservation)

Coated Wells	96 holes	Enzyme Conjugate	12ml
10Ã— specimen dilution (Sample Buffer)	12ml	20Ã—Wash Buffer	50ml
Standards: 40ng / 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. Collection of specimens: serum, plasma (EDTA, citrate, heparin anticoagulation), cell culture supernatant, tissue homogenate, etc., as early as possible, stored at 2-8 Â° C for 48 hours; longer time must be frozen (-20 Â° C Or -70 Â°C) to avoid repeated freezing and thawing. Serum and plasma were diluted 1 : 2 (take 100 ul, add 100 ul of standard dilution, dilute 2 times). The cell culture supernatant can be directly detected without dilution.

2. Standard solution preparation: Add 1 ml of distilled water before use and mix well to prepare 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 10Ã— 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 Â°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 Â° 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 Â° 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 Â° 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 â€‹â€‹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 CCR-1 content on the graph based on the OD value of the sample, and multiply by the dilution factor.

Kit performance

1. Sensitivity: The minimum CCR-1 detection concentration is less than 40pg/ml.

2. Specificity: Recombinant or natural human CCR-1 can be detected simultaneously. Does not cross-react with other human cytokines.

3. Repeatability: The coefficient of variation in the plate and plate is less than 12%.

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!

Fruit Powder

Fruit Powder

Fruit powder is made from 6-7 mature and fig or fig dried fruit by raw material processing, squeezing juice, filtering, clarifying, concentrating, spray drying, cooling, packaging and other steps.

Key points of production process:

Selection of raw materials: select 6-7 mature and fresh figs or dried figs. Raw material treatment and pressing for juice: wash the raw materials, add materials according to the proportion of 1kg water plus 1kg fruit, or 1kg dried fruit plus 5kg water, put them into a stainless steel pot, heat them to 85-90 â„ƒ, keep them for 20-30 minutes, then stop heating, stand for 24 hours, and press for juice. Filtration and clarification: it is filtered by a screen filter, and then natural clarification or enzyme clarification. concentrate; Atmospheric pressure concentration and vacuum concentration can be adopted. It is concentrated under normal pressure in a stainless steel double-layer pot, and the heating steam pressure is 2.5kg/cm2. In the concentration process, pay attention to stirring, accelerate water evaporation, prevent coking, and concentrate to make the solid reach 28%. Each concentration should not be fed too much, and the time should be 40 minutes. Vacuum concentration, concentration under reduced pressure and lower temperature. Heating, steam pressure 1.5kg/cm2, temperature 50 â„ƒ.

Spray drying: high pressure spray equipment was used to spray the dried fig juice concentrate. The feeding temperature was 50-60 C, the working pressure of the high pressure pump was 180 kg / cm, the dry additive paste powder was added 0.5%, the air inlet temperature was 120 C, and the outlet temperature was 75-78 C. Cooling and packaging: the dried fig powder is cooled quickly, and then packaged and sealed.

For Example: Blueberry Juice Powder

In addition to conventional sugar, acid and VC, blueberry fruit is rich in VE, VA, VB, SOD, arbutin, protein, anthocyanin, edible fiber and mineral elements such as K, Fe, Zn and ca. According to the analysis and determination of 14 varieties of blueberry fruits introduced from the United States, the content of anthocyanin pigment per 100 grams of blueberry fresh fruits is as high as 163 mg, protein 400-700 mg, fat 500-600 mg, carbohydrate 12.3-15.3 mg, vitamin A as high as 81-100 international units, vitamin E 2.7-9.5 micrograms and sod5.39 international units. Vitamins are higher than other fruits. Trace elements are also high, with 220-920 micrograms of calcium, 98-274 micrograms of phosphorus, 114-249 micrograms of magnesium, 2.1-4.3 micrograms of zinc, 7.6-30.0 micrograms of iron, 0.8-1.2 micrograms of germanium and 2.0-3.2 micrograms of copper per gram of fresh fruit.

It is precisely because blueberry fruit is rich in nutrients. It is a nutritional and health fruit with high amino acids, high zinc, high calcium, high iron, high copper and high vitamins. It not only has good nutrition and health care function, but also has the functions of preventing brain nerve aging, strengthening heart, anti-cancer, softening blood vessels, enhancing human immunity and so on.

Health care function

Blueberries can delay memory decline and prevent heart disease, so they are regarded as super fruits. The US daily health news reported that recent research has added a good reputation to super fruit. Eating more blueberries or drinking blueberry juice can help prevent colon cancer.

The study found that blueberries can play this role because blueberries and other fruits contain a natural compound called pterostilbene, which helps prevent pre cancer damage to the body. Pterostilbene is an antioxidant and anti-inflammatory agent, which is found in blueberries and blackberries.

Blueberries contain a lot of purple components, anthocyanins.

Blueberry fruit can significantly enhance vision and eliminate eye fatigue; Nourish skin; Delaying brain nerve aging; It has therapeutic effect on capillary disease caused by diabetes. Enhance cardiac function; Prevention of Alzheimer's disease and many other medicinal values.

Anthocyanins in blueberry fruit are very powerful antioxidants, which can help prevent the formation of plaque in arteries and a variety of cancers (folic acid prevents cervical cancer, etc.), reduce the possibility of cancer, reduce heart disease and delay aging.

blueberry powder

Fruit powder, Fruit Juice Powder, Watermelon Juice Powder

Xi'an Tian Guangyuan Biotech Co., Ltd. , https://www.tgybiotech.com