Rat tumor necrosis factor beta (TNF - β) detection kit

Specifications96T/48T

Product UsageFor scientific research testing only, not for clinical use

Company Services:ELISAtest kit Free proxy testing and inspection

Types of test samples: serum, plasma, urine, tissue homogenate, cell supernatant, lavage fluid, feces, etc

LiBo BiologyThe supplied species include: rats, mice, guinea pigs, rabbits, dogs, monkeys, pigs, cows, sheep, chickens, ducks, geese, plants, fish, insect ELISA kits, etc

Rat tumor necrosis factor beta (TNF - β) detection kit

Experimental principle of rat tumor necrosis factor beta (TNF - β) assay kit：
The rat tumor necrosis factor beta (TNF - β) assay kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). Standard samples with known concentrations of the test substance and samples with unknown concentrations are added to a microporous ELISA plate for detection. Firstly, incubate the test substance and biotinylated antibody simultaneously. After washing, add HRP labeled with avidin. After further incubation and washing, unbound enzyme complexes are removed, and then substrates A and B are added to interact with the enzyme complexes simultaneously. Generate color. The depth of color is proportional to the concentration of the substance to be tested in the sample.

Rat tumor necrosis factor beta (TNF - β)Kit Content and Preparation

Rat tumor necrosis factor beta (TNF - β) test kit with self provided materials
1) Distilled water.
2）加样器： 5ul、10ul、50ul、100ul、200ul、500ul、1000ul。
3) Oscillators and magnetic stirrers, etc.

Safety of rat tumor necrosis factor beta (TNF - β) assay kit
1) Avoid direct contact with termination solution and substrates A and B. Once in contact with these liquids, please rinse with water as soon as possible.
2) Do not eat, drink, smoke or use cosmetics during the experiment.
3) Do not use your mouth to suck any ingredients from the reagent kit.

Precautions for operating the rat tumor necrosis factor beta (TNF - β) kit
1) Reagents should be stored according to the label instructions and returned to room temperature before use. The diluted standard should be discarded and cannot be stored.
2) The Flat noodles not used in the experiment shall be immediately put back into the packaging bag and sealed for storage to avoid deterioration.
3) Other unused reagents should be packaged or covered. Do not mix reagents of different batches. Use before shelf life.
4) Use disposable suction tips to avoid cross contamination, and avoid using sample dispensers with metal parts when suctioning termination solution and substrates A and B.
5) Use clean plastic containers to prepare detergent. Mix all components and samples in the reagent kit thoroughly before use.
6) When washing the enzyme-linked immunosorbent assay (ELISA) plate, it should be thoroughly dried. Do not directly put the absorbent paper into the ELISA reaction well to absorb water.
7) Substrate A should evaporate and avoid opening the lid for a long time. Substrate B is sensitive to light and should be avoided from prolonged exposure to light. Avoid contact with hands, toxic. After the experiment is completed, the OD value should be read immediately.
8) The order of adding reagents should be consistent to ensure that the incubation time for all reaction plate wells is the same.
9) Perform incubation operations according to the time, amount, and sequence indicated in the instructions.

Collection, processing, and storage methods of rat tumor necrosis factor beta (TNF - β) assay kit samples
1) Serum - Avoid any cellular stimulation during operation. Use test tubes without pyrogen and endotoxin. After collecting blood, centrifuge at 1000 × g for 10 minutes to quickly and carefully separate serum and red blood cells.
2) Plasma - EDTA, citrate, heparin plasma can be used for detection. Centrifuge at 1000 × g for 30 minutes to remove particles.
3) Cell supernatant - Centrifuge at 1000 × g for 10 minutes to remove particles and polymers.
4) Tissue homogenate - add an appropriate amount of physiological saline and crush the tissue. Centrifuge at 1000 × g for 10 minutes and collect the supernatant
5) Storage - If the sample is not used immediately, it should be divided into small parts and stored at -70 ℃ to avoid repeated freezing. Try not to use hemolytic or hyperlipidemic blood as much as possible. If there are a large number of particles in the serum, centrifuge or filter before detection. Do not thaw by heating at 37 ℃ or higher. Thaw at room temperature and ensure that the sample is evenly thawed.

Preparation of rat tumor necrosis factor beta (TNF - β) assay kit reagents
1) Standard: The series dilution of standard should be prepared during the experiment and cannot be stored. Shake and mix the standard before dilution.
2) Dilution of washing buffer (50x): Dilute 50 times with distilled water.

Operation steps of rat tumor necrosis factor beta (TNF - β) kit
1) Before use, mix all reagents thoroughly. Do not make the liquid produce a lot of foam, so as not to add a lot of bubbles when adding samples, which will cause errors in adding samples.
2) Determine the number of Flat noodles required according to the number of samples to be tested plus the number of standards. It is recommended to make duplicate holes for each standard and blank hole. Each sample is determined according to its own quantity, and those that can be reused should be reused as much as possible. Dilute the specimen with a 1:1 dilution solution and add 50ul to the reaction well.
3) Add 50ul of diluted standard substance to the reaction well and 50ul of the test sample to the reaction well. Immediately add 50ul of biotinylated antibody. Cover the membrane plate, gently shake and mix well, and incubate at 37 ℃ for 1 hour.
4) Shake off the liquid in the holes, fill each hole with detergent, shake for 30 seconds, shake off the detergent, and pat dry with absorbent paper. Repeat this operation 3 times. If washing with a washing machine, increase the number of washes by one.
5) Add 80ul of affinity chain enzyme HRP to each well, gently shake and mix well, and incubate at 37 ℃ for 30 minutes.
6) Shake off the liquid in the holes, fill each hole with detergent, shake for 30 seconds, shake off the detergent, and pat dry with absorbent paper. Repeat this operation 3 times. If washing with a washing machine, increase the number of washes by one.
7) Add 50ul of substrate A and B to each well, gently shake and mix well, and incubate at 37 ℃ for 10 minutes. Avoid exposure to light.
8) Take out the enzyme-linked immunosorbent assay (ELISA) plate and quickly add 50ul of termination solution. The results should be measured immediately after adding the termination solution.
9) Measure the OD values of each well at a wavelength of 450nm.