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Human Fatty Acid Binding Protein (FABP) Enzyme Linked Immunoassay (ELISA)
Kit instruction manual
This reagent is for research use only
Purpose: This kit is used to determine the content of fatty acid binding protein (FABP) in human serum, plasma and related liquid samples.
Kit composition:
Kit composition
48 hole configuration
96-well configuration
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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 ℃
Standard product: 2700ng / L
0.5ml × 1 bottle
0.5ml × 1 bottle
Store at 2-8 ℃
Standard dilution
1.5ml × 1 bottle
1.5ml × 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 ℃
Experimental principle:
This kit uses the double antibody sandwich method to determine the level of human fatty acid binding protein (FABP) in the specimen. Microporous plates were coated with purified human fatty acid binding protein (FABP) antibody to make solid phase antibodies. Fatty acid binding protein (FABP) was added to the microwells coated with mAb in turn, followed by HRP labeled fatty acid binding protein (FABP) ) The antibody binds to form an antibody-antigen-enzyme-labeled antibody complex. 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 color depth is positively correlated with the fatty acid binding protein (FABP) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human fatty acid binding protein (FABP) in the sample was calculated by a standard curve.
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. 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. Dilution and loading of standard products: set 10 standard wells on the enzyme-coated plate, add 100 μl of the standard products in the first and second wells, and then add the standard products in the first and second wells. 50μl of diluent, mix well; then take 100μl from the first well and the second well and add them to the third and fourth wells respectively, and then add 50μl of standard diluent to the third and fourth wells respectively, mix well; Then take 50μl each in the third and fourth wells and discard it, then add 50μl each to the fifth and sixth wells, and then add 50ul of the standard dilution solution to the fifth and sixth wells respectively, and mix well; After mixing, take 50μl from the fifth and sixth wells and add them to the seventh and eighth wells respectively. Then add 50μl of the standard dilution solution to the seventh and eighth wells respectively. Take 50μl from the eight wells and add them to the ninth and tenth wells. Then add 50μl of the standard dilution solution to the ninth and tenth wells. After mixing, take 50μl from the ninth and tenth wells and discard. (After dilution, the volume of each well is 50μl, and the concentration is 1800ng / L, 1200ng / L, 600ng / L, 300ng / L, 150ng / L)
2. Add samples: set up blank wells (the blank control wells do not add samples and enzyme reagents, the rest of the steps are the same) and the sample wells to be tested. Add 40μl of sample diluent to the test sample well of the enzyme-coated plate, and then add 10μl of the test sample (the final dilution of the sample is 5 times). Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix.
3. Incubation: Seal the plate with a sealing plate and incubate at 37 ° C for 30 minutes.
4. Mixing solution: Dilute 30 times (20 times of 48T) concentrated washing liquid with distilled water 30 times (20 times of 48T) and then use.
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 5 times and 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, then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes.
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.
Precautions:
1. 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.
2. 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.
3. The sampler should be used at each step of sample addition, and the accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples.
4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the test substance in the specimen is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please first dilute it with a certain multiple (n times) of the sample diluent and then determine it. When calculating, please multiply the total dilution Multiple (× n × 5).
5. The sealing film is limited to one-time use to avoid cross-contamination.
6. Please keep the substrate away from light.
7. Strictly follow the instructions, and the test results must be determined by the microplate reader.
8. All samples, washing liquids and various wastes should be treated as infectious agents.
9. The components of different batches of this reagent shall not be mixed.
10. If there is any difference with the English manual, the English manual shall prevail.
Kit performance:
1. The correlation coefficient R between the linear regression of the sample and the expected concentration is above 0.95.
2. The batch and approval shall be less than 9% and 11% respectively
examination range:
100ng / L -2000ng / L
Calculation:
Taking the concentration of the standard as the abscissa and the OD value as the ordinate,
Draw a standard curve on coordinate paper, according to the OD of the sample
The value is determined by the standard curve; then multiplied by the dilution
Multiple; or calculate the standard using the concentration and OD value of the standard
The linear regression equation of the quasi-curve, the OD value of the sample
Substitute into the equation, calculate the sample concentration, and multiply by the dilution
The multiple is the actual concentration of the sample.
Storage conditions and validity period:
1. Store the kit: 2-8 ℃.
2. Validity: 6 months
Fecal Suction truck
Suction truck (Fecal suction truck), mainly applied to the lower septic tanks, sewer and gutter cleaning and other environmental clean-up of special vehicles, and sprinkler, garbage trucks, and known as the three sanitation models.
Fecal Suction truck
Brief introduction
Suction truck main vehicle chassis, tanks, fecal suction devices and other components. Furui Ka models commonly used Dongfeng, Dongfeng DLK, 140, 145, after the double bridge, top four after eight other models.
Fecal Suction truck consists of: oil and water separator, moisture separator, special vacuum suction pump manure, volume gauge, pipe network system, suction catheter material, gravity valve, vacuum tank, connected (depending on the dung window), automatic anti overflow valve.
Scope
Mainly for urban and rural sanitation sector suction and delivery of manure and other waste water. Suitable for pumping manure, sewage, sludge and slurry mixed with small debris suspended in the liquid, used in municipal sanitation, small medium and large industrial and mining enterprises, residential, schools, transport septic tank cleaning, pipeline cleaning dredge city, marsh plant biogas and other clean-up liquid.
Optional models
1, pay attention to the quality of plates, vacuum tightness, poor materials likely to cause deformation of the tank and other issues.
2, vacuum package selection, bearing capacity and temperature difference of perception. Domestic kit devaluation Veyron series.
3, gravity valve selection, according to the local operating environment, temperature and other options.
Product Categories
By brand
CLW Fecal suction truck, Dongfeng Fecal suction truck, FAW suction truck, FOTON suction truck, JAC suction truck, sino truck suction truck, ISUZU suction truck.
In accordance with the shape
Single bridge suction truck, suction truck Shuangqiao, flat suction truck, suction truck tip
According to Variety
4000L Fecal suction truck, 6000L suction truck, 8000L suction truck, 10000L suction truck 12000L suction truck,15000L suction truck .
Use according to
Tipper suction truck, three suction truck (agricultural suction truck)
User's Guide
Job preparation
(1) suction truck as close to the rear of the operating point, parking.
(2) will return the tank to pull the handle straight cocks rotation axis of the intake pipe about 45 ° angle of observation into the pipeline, the lubricating oil should flow.
(3) take the box-side door open, remove the fecal suction hose, making it swing back, without bending phenomenon.
Sucking operation
(1). The fecal suction hose as won pumped liquid manure to ensure that the pipe end during the job and always below 300mm from the liquid surface.
(2) would be pushed to the four-way valve handle perpendicular to the ground.
(3) would be linked to the transmission into neutral and start the engine, disengage the clutch, the power take-gear switch is pulled back that is to take power, the vacuum pump starts running.
(4) Open cab toolbox Xiangqi pull the switch, turn on the power.
(5) The operator should pay attention to monitoring the situation near the suction manure hose imports, with the appropriate tools to block debris to prevent blockage.
(6) After the viewer through the operator head with the Ministry, when the liquid level reaches the middle of the mirror to observe shall inform the driver, colleagues shall promptly fecal suction hose pulled from the ground or off the four-way valve. Under normal circumstances, this time Xiangqi audible and visual signals, the driver receives a signal should reduce the throttle, the power take off switch forward shirk stall, pump stopped moving forward, according to the Xiangqi switch off the power .
(7) The fuel tank straight cocks rotation axis parallel to the shank plate and into the mailbox is closed.
(8) After rinsing hose, put it back down station box, shut the side door, and above the boom toward the cab.
(8) close the spill valve, so that it can handle the road perpendicular to the axis.
(9) the suction truck left the operating point.
Unloading row
(1) toward the suction manure hose storage within the bog.
(2) the four-way valve handle to pull parallel to the ground, open the spill valve, so that it is parallel to the axis of the handle to the pipe.
(3) linked to the transmission into neutral and start the engine, disengage the clutch, the power take-gear switch is pulled back that is to take power, the vacuum pump starts running.
(4) inside the tank liquid manure discharge unloaded, the driver should be promptly handles power take off the file that is pushed forward, the vacuum pump is stopped.
(5) The fuel tank straight cocks rotation axis parallel to the shank plate and into a mailbox that is closed, after rinsing hose, put it back down station box, shut the side door, and above the boom toward the cab.
(6) the suction truck left the operating point.
Fecal Suction Truck,Vacuum Fecal Suction Truck,Fecal Suction Tanker Truck,Sewage Fecal Suction Truck
CLW GROUP TRUCK , https://www.clwgrouptruck.net