The principle of human â…¢ collagen (Col â…¢) ELISA experiment
English name: Human Collagen Type â…¢, Col â…¢ ELISA
1. The level of human type â…¢ collagen (Col â…¢) ELISAKit was determined by double antibody sandwich method.
2. First coat the microplate with purified human type III collagen (Col III) ELISAKit antibody to make a solid phase antibody.
3. Then add collagen â…¢ (Col â…¢) ELISAKit to the microwells coated with mAb in turn, and then combine with HRP-labeled collagen â…¢ (Col â…¢) ELISAKit antibody to form an antibody-antigen-enzyme label antibody complex After thorough washing, add substrate TMB to develop color. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid.
4. The color depth is positively correlated with the type â…¢ collagen (Col â…¢) ELISAKit in the sample.
5. Finally, the absorbance (OD value) is measured with a microplate reader at a wavelength of 450 nm, and the type â…¢ collagen (Col â…¢) ELISA Kit in the sample is calculated by the standard curve.
Objective: To determine the level of type â…¢ collagen (Col â…¢) ELISAKit in human serum, plasma and related liquid samples.
6. Serum: Blood coagulates naturally at room temperature for 10-20 minutes, centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully and centrifuge again if a precipitate appears during storage. Avoid any cell stimulation during the operation. Use test tubes free of pyrogens and endotoxins.
7. Plasma: EDTA or sodium citrate should be selected as the anticoagulant according to the requirements of the specimen. After mixing for 10-20 minutes, centrifuge for about 20 minutes (2000-3000 rpm) to remove particles. Collect the supernatant carefully, and centrifuge again if there is any precipitate.
safety:
1. Avoid direct contact with stop solution and substrate. If you accidentally come into contact with these liquids, please rinse them with water as soon as possible.
2. Do not eat, drink, smoke or use cosmetics during the experiment.
3. Do not touch any ingredients in the kit with your mouth.
4 Keep Elisa kit away from children
Bring your own items:
1. 37 ℃ incubator
2. Standard specification microplate reader
3. Precision pipette and disposable tip
4. Distilled water
5. Disposable test tube
6. Absorbent paper
Storage conditions and validity period:
1. Kit storage :; 2-8 ℃.
2. Validity: 6 months
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