Manual of Schistosoma ELISA Kit
This reagent is for research purpose only: This kit is used to detect the level of Schistosoma in human serum, plasma and related liquid samples, and hematological diagnosis of infected people.
Experimental principle:
This kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to determine human Schistosoma (Schistosoma) in the specimen. Microporous plates are coated with purified Schistosoma antibody to make solid-phase antibodies, which can be combined with Schistosoma (Schistosoma) in the sample. After washing to remove unbound antibodies and other components, they are then labeled with HRP schistosome Schistosoma) antibodies bind to form an antibody-antigen-enzyme-labeled antibody complex, and after thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm and compared with the CUTOFF value to determine the presence or absence of Schistosoma in the specimen.
Kit composition:
Kit composition
48 hole configuration
96-well configuration
save
Instructions
1 serving
1 serving
Sealing film
2 pieces (48)
2 pieces (96)
sealed bag
1
1
Enzyme coated plate
1 × 48
1 × 96
Store at 2-8 ℃
Negative control
0.5ml × 1 bottle
0.5ml × 1 bottle
Store at 2-8 ℃
Positive control
0.5ml × 1 bottle
0.5ml × 1 bottle
Store at 2-8 ℃
Enzyme reagent
3 ml × 1 bottle
6 ml × 1 bottle
Store at 2-8 ℃
Sample diluent
3 ml × 1 bottle
6 ml × 1 bottle
Store at 2-8 ℃
Developer A liquid
3 ml × 1 bottle
6 ml × 1 bottle
Store at 2-8 ℃
Developer B liquid
3 ml × 1 bottle
6 ml × 1 bottle
Store at 2-8 ℃
Stop solution
3ml × 1 bottle
6ml × 1 bottle
Store at 2-8 ℃
Concentrated washing liquid
(20ml × 20 times) × 1 bottle
(20ml × 30 times) × 1 bottle
Store at 2-8 ℃
Sample processing and requirements:
1. Serum: room temperature blood coagulates naturally 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.
2. Plasma: EDTA or sodium citrate should be selected as the anticoagulant according to the requirements of the specimen, mixed for 10-20 minutes, and centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
3. Urine: collected in a sterile tube and centrifuged for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, centrifuge again. Pleural and ascites, cerebrospinal fluid reference implementation.
4. Cell culture supernatant: When detecting secreted components, collect with a sterile tube. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. When detecting the components inside the cells, dilute the cell suspension with PBS (PH7.2-7.4), and the cell concentration will reach about 1 million / ml. Through repeated freezing and thawing, the cells are destroyed and the intracellular components are released. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. If a precipitate forms during storage, it should be centrifuged again.
5. Organize the specimen: after cutting the specimen, weigh it. Add a certain amount of PBS, PH7.4. Quickly freeze and save with liquid nitrogen for later use. After the specimen melts, it still maintains a temperature of 2-8 ° C. Add a certain amount of PBS (PH7.4) and homogenize the specimen with a manual or homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. After aliquoting, a portion is to be tested, and the rest is frozen for future use.
6. The specimen should be extracted as soon as possible after collection. The extraction should be carried out according to relevant literature. The experiment should be carried out as soon as possible after extraction. If the test cannot be performed immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided.
7. The sample containing NaN3 cannot be detected because NaN3 inhibits the activity of horseradish peroxidase (HRP).
Steps:
1. Numbering: serially number the corresponding microwells of the sample, each plate should be set with 2 wells for negative control, 2 wells for positive control, and 1 well for blank control (the blank control well does not add sample and enzyme reagent, the rest of the steps are the same)
2. Add sample: add negative control and positive control 50μl to the negative and positive control wells respectively. Then add 40μl of sample diluent to the sample well, and then add 10μl of the sample to be tested. Add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, gently shake to mix,
3. Incubation: Seal the plate with a sealing plate and incubate at 37 ° C for 30 minutes.
4. Mixing solution: Add 30 times (20 times of 48T) concentrated washing liquid to 600ml with distilled water and set aside
5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing liquid, let it stand for 30 seconds and then discard, repeat this 5 times, pat dry.
6. Add enzyme: add 50μl of enzyme label reagent to each well, except blank well.
7. Incubation: The operation is the same as 3.
8. Washing: The operation is the same as 5.
9. Color development: add 50μl of developer A to each well, and then add 50μl of developer B, mix gently, and develop for 15 minutes in the dark at 37 ℃
10. Termination: Add 50μl of stop solution to each well to stop the reaction (at this time the blue will turn to yellow).
11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution.
Result judgment:
Test effectiveness: the average of positive control wells ≥1.00; the average of negative control wells ≤0.10
Calculation of critical value (CUT OFF): critical value = average value of negative control wells + 0.15
Negative judgment: the sample with OD value <cut-off value (CUT OFF) is negative for Schistosoma Ab (Schistosoma Ab)
Positive judgment: the sample with OD value ≥ critical value (CUT OFF) is positive for human Schistosoma Ab (Schistosoma Ab)
Precautions
1. The operation is strictly in accordance with the instructions. The components of different batches of this reagent must not be mixed.
2. The kit should be equilibrated at room temperature for 15-30 minutes before being taken out of the refrigerated environment. If the enzyme label coated plate is unopened, the strip should be stored in a sealed bag.
3. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in a water bath during dilution, and the results will not be affected during washing.
4. The sealing film is limited to one-time use to avoid cross-contamination.
5. Please keep the substrate away from light.
6. The test results must be determined based on the reading of the microplate reader. When using dual wavelength detection, the reference wavelength is 630nm
7. All samples, washing liquids and various wastes should be treated as infectious agents. The stop solution is 2M sulfuric acid, and you must pay attention to safety when using it.
Storage conditions and validity period
1. Kit storage :; 2-8 ℃.
2. Validity: 6 months
Xiamen Huijia Biotechnology Co., Ltd. is a professional agent for many well-known brands in the international life science field. Committed to the sales and promotion of various ELISA kits, immunohistochemical kits, primary and secondary antibodies, cytokines, biological reagents, pipettes, and consumables.
Office chairs can be used in companies, homes, and various commercial places. Our office chairs are comfortable to sit on, with many colors and varieties. The backrest is made of mesh cloth, and the cushion is made of sponge, breathable, dirt-resistant, suitable for office And games.Myopia in children is mostly caused by bad habits in the learning process, such as improper sitting posture and poor lighting. The learning environment created by a good learning table is still helpful for preventing myopia, so buy anti-myopia learning desks for children. There is still a certain effect, let children develop good study habits, which will accompany them for a lifetime.
After the child goes to school, they can use the study table. The study table on the market can adjust the height of the table according to the height of the child. The study table is an essential tool for children to do homework. For example, there is a great and handsome study table on the market. The function is intelligent lifting and lowering, and the desktop can be folded. , When a person sits in front of the table, they naturally enter the concave table top, and the two arms droop naturally. Because the two elbows are supported by the concave plate, the body naturally does not lean forward, which effectively solves the shortcomings of lying on the stomach to write and study. When shopping for a Study Desk, be sure to keep these must-have features in mind. The focus of the study table is to correct the sitting posture and prevent myopia, which is a foldable study table with a concave table top.
Pu Leather Office Chair,Office Chair,Nordic Office Chair,Stylish Office Chair
Igrow Technology Co.,LTD , https://www.igrowdesks.com