Rat (Rat) 1-sphingosine 1 (S1P) ELISA Kit

The kit can only be used for scientific research and should not be used for medical diagnosis.

Rat (Rat) 1-sphingosine 1 (S1P) ELISA Kit

Detection principle

The kit uses a double antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). To the coated microwells previously coated with the sphingosine 1 -phosphate (S1P) antibody, the specimen, the standard, and the HRP-labeled detection antibody were sequentially added, and the cells were washed and thoroughly washed. Using the substrate TMB to develop color, TMB is converted to blue under the catalysis of peroxidase and converted to the final yellow color by the action of an acid. The color depth is positively correlated with sphingosine-1-phosphate (S1P) in the sample. The absorbance (OD value) was measured at 450 nm using a microplate reader to calculate the sample concentration.

Sample collection, processing and storage methods

1. Serum: Use a tube containing no pyrogen and endotoxin. Avoid any cell irritation during the procedure. After collecting the blood, centrifuge and centrifuge for 10 minutes at 3000 rpm to quickly and carefully separate the serum and red blood cells.

2. Plasma: EDTA, citrate or heparin anticoagulation. The supernatant was taken by centrifugation at 3000 rpm for 30 minutes.

3. Cell supernatant: Centrifuge at 3000 rpm for 10 minutes to remove particles and polymer.

4. Tissue homogenization: The tissue is mashed by adding appropriate amount of physiological saline. The supernatant was taken by centrifugation at 3000 rpm for 10 minutes.

5. Storage: If the sample is not detected in time after collection, please dispense it once, freeze it at -20 °C, avoid repeated freezing and thawing, thaw at room temperature and ensure that the sample is fully thawed evenly.

Bring your own items

1. Microplate reader (450nm)
2. High-precision sampler and tip: 0.5-10uL, 2-20uL, 20-200uL, 200-1000uL
3. 37 ° C incubator

Operational precautions

1. The kit was stored at 2-8 ° C and equilibrated for 20 minutes at room temperature before use. The concentrated washing liquid taken out from the refrigerator will crystallize, which is a normal phenomenon, and the water bath is heated to completely dissolve the crystals before use.
2. The slats not used in the experiment should be immediately put back into the ziplock bag and sealed (low temperature dry) for storage.
3. The S0 standard with a concentration of 0 can be regarded as a negative control or blank; the sample has been diluted 5 times according to the instructions, and the final result multiplied by 5 is the actual concentration of the sample.
4. Incubation is carried out in strict accordance with the time indicated in the instructions, the amount of liquid added and the order.
5. Shake well all liquid components before use.

Kit composition

Note: The concentration of standard (S0-S5) is: 0, 50, 100, 200, 400, 800 nmol/L

Reagent preparation

Dilution of 20× Wash Buffer: Distilled water was diluted 1:20, ie 1 part of 20× Wash Buffer plus 19 parts of distilled water.

Washing method

1. Manually wash the plate: Drain the liquid in the hole, fill each hole with the washing liquid, let stand for 1 min, then drain the liquid in the hole, pat dry on the absorbent paper, and wash the plate 5 times.
2. Automatic washing machine: Inject 350 μL of washing solution into each well, soak for 1 min, and wash the plate 5 times.

Steps

1. Remove the required slats from the aluminum foil bag after 20 minutes of equilibration at room temperature, and seal the remaining slats back to 4 °C with a ziplock bag.

2. Set standard and sample wells, and add standard concentration of 50 μL to each standard.

3. The sample well is first added with 10 μL of the sample to be tested, and then the sample dilution is 40 μL; the blank well is not added.

4. In addition to the blank wells, add 100 μL of horseradish peroxidase (HRP)-labeled detection antibody to each well of the standard wells and sample wells, seal the wells with a sealing membrane, and incubate in a 37 ° C water bath or incubator. 60min.

5. Discard the liquid, pat dry on the absorbent paper, fill each well with the washing solution, let stand for 1 min, remove the washing solution, pat dry on the absorbent paper, and repeat the washing 5 times (can also be washed with a washing machine).

6. Add 50 μL of substrate A and B to each well and incubate at 37 ° C for 15 min in the dark.

7. Add 50 μL of stop solution to each well, and measure the OD value of each well at a wavelength of 450 nm within 15 min.

Result judgment

Draw a standard curve: In the Excel worksheet, the standard concentration is used as the abscissa, and the OD value is plotted as the ordinate. The linear regression curve of the standard is drawn, and the concentration values ​​of each sample are calculated according to the curve equation.

Kit performance

1. Accuracy: The linear regression coefficient of the standard and the expected concentration correlation coefficient R value, greater than or equal to 0.9900.
2. Sensitivity: The minimum detection concentration is less than 1.0 nmol/L.
3. Specificity: Does not cross-react with other soluble structural analogs.
4. Repeatability: The coefficient of variation between the plates and the plates is less than 15%.
5. Storage: 2-8 ° C, protected from light and moisture.
6. Validity: 6 months

Disclaimer

1. The kit is for research use only and should not be used for clinical trials or human experiments. Otherwise, the consequences will be borne by the experimenter and the company will not be responsible.
2. In strict accordance with the instructions, the experimenter violates the instructions, and the consequences are borne by the experimenter.

Shanghai Yuduo Biotechnology Co., Ltd. produces and sells elisa kit, animal serum, plasma, whole blood, antigen and antibody, gold standard kit test card, antiserum, biological reagents, culture medium, laboratory equipment consumables, chemical reagents, biological products. , standard products, reference products, biochemical immunological products, immunoaffinity columns, strains, plasmids, indoor quality control products, cells, ice packs, agent imported products, etc.

Large favorably! Welcome dealers to discuss cooperation matters!

Contact Phone Contact QQ

Cosmetic Ingredients

Cosmetics are compound mixtures made from various raw materials after reasonable blending and processing. There are many types of cosmetics raw materials with different properties. According to the raw material properties and uses of cosmetics, it can be roughly divided into two categories: base raw materials and auxiliary raw materials. The former is a main raw material of cosmetics, which occupies a large proportion in cosmetic formulations, and is the main functional substance in cosmetics. The latter is responsible for shaping, stabilizing or imparting color, fragrance and other characteristics to cosmetics. These substances are not used in large amounts in cosmetic formulations, but are extremely important. Cosmetics are chemical mixtures made of natural, synthetic or extracted substances with different functions as raw materials and processed through production processes such as heating, stirring and emulsification.

Collagen peptide, Arbutin, Coenzyme Q10, GSH, Cosmetic Raw Materials, lipoic acid

Xi'an Gawen Biotechnology Co., Ltd , https://www.ahualyn-bios.com