Mouse HYD ELISA Test Kit Instruction Manual
Test principle:
The HYD kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards of known HYD concentrations and samples of unknown concentration are added to the microplates for detection. HYD and biotinylated antibodies were first incubated simultaneously. After washing, avidin-labeled HRP was added. After incubation and washing, the unbound enzyme conjugate is removed, then the substrates A, B, and the enzyme conjugate are added simultaneously. Produce color. The depth of the color is proportional to the concentration of HYD in the sample.
Content and its preparation:
Kit component 96-well configuration 48-well configuration
96/48 person ELISA plate 1 plate (96T) half plate (48T)
Plastic diaphragm cover 1 block
Standard : 240ng/ml 1 bottle (1.0ml) 1 bottle (0.5ml)
Blank control 1 bottle (1.0ml) 1 bottle (0.5ml)
Standard dilution buffer 1 bottle (8.0ml) 1 bottle (4.0ml)
Biotin-labeled anti-HYD antibody 1 bottle (8.0ml) 1 bottle (4.0ml)
Affinity Chain Enzyme-HRP 1 Bottle (12ml) 1 Bottle (5ml)
1 bottle of washing buffer (20ml) 1 bottle (10ml)
Substrate A 1 bottle (6.0ml) 1 bottle (3.0ml)
Substrate B 1 bottle (6.0ml) 1 bottle (3.0ml)
Stop solution 1 bottle (6.0ml) 1 bottle (3.0ml)
Bring your own materials:
1. Distilled water.
2. Sampler: 5ul, 10ul, 50ul, 100ul, 200ul, 500ul, 1000ul.
3. Oscillators and magnetic stirrers.
4.
Sample collection, processing and storage methods:
1. Serum... Avoid any cell irritation during the procedure. Use tubes without pyrogens and endotoxins. After collecting blood, centrifuge at 1000 × g for 10 minutes to red blood
The cells are quickly and carefully separated.
2, plasma ... EDTA, citrate, heparin plasma can be used for testing. The pellet was removed by centrifugation at 1000 x g for 30 minutes.
3. Cell supernatant. Centrifuge at 1000 xg for 10 minutes to remove particles and polymer.
4. Tissue homogenization... Add the appropriate amount of normal saline to the tissue. Centrifuge at 1000 xg for 10 minutes and take the supernatant.
5. Save... If the sample is not used immediately, it should be divided into small parts - 70 ° C to avoid repeated freezing. Do not use hemolysis or hyperlipemia as much as possible. If there are large amounts of particles in the serum, centrifuge or filter before testing. Do not heat thaw at 37 ° C or higher. It should be thawed at room temperature and ensure that the sample is fully thawed evenly.
Operational notes:
â— Reagents should be stored according to the label instructions and returned to room temperature before use. Standards after dilution should be discarded and cannot be stored.
â— The slats not used in the experiment should be immediately put back into the bag and sealed to prevent deterioration.
â— Other reagents that are not used should be packaged or covered. Do not mix reagents of different batches. Use before warranty.
â— Use disposable tips to avoid cross-contamination. Avoid pipettes with metal parts when drawing stop solution and substrate A and B.
â— Use a clean plastic container to configure the wash solution. Mix all the ingredients and samples in the kit thoroughly before use.
â— Substrate A should be volatilized to avoid opening the lid for a long time. Substrate B is sensitive to light and avoids prolonged exposure to light. Avoid contact with hands and be toxic. The OD value should be read immediately after the experiment is completed.
â— The order of adding reagents should be the same to ensure that all wells are incubated for the same time.
â— Carry out the incubation according to the time indicated in the instruction manual, the amount of liquid addition and the order.
safety:
1. Avoid direct contact with the stop solution and substrate A, B. Once exposed to these liquids, rinse with water as soon as possible.
2. Do not eat, drink, smoke or use cosmetics.
3. Do not use the mouth to absorb any ingredients in the kit.
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