Mouse complement 1 inhibitor antibody (GC1INH) ELISA kit instruction manual
This reagent is for research use only. Specimen: serum or plasma
This reagent is for research use only. Purpose: This kit is used to determine the content of α-galactosyl antibody (GC1INH) in human serum, plasma and related liquid samples.
Test principle:
The GC1INH kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards of GC1INH concentration and samples of unknown concentration are added to the microplates for detection. GC1INH and biotinylated antibodies were first incubated simultaneously. After washing with the mouse complement 1 inhibitor antibody, 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 GC1INH in the sample.
Kit composition:
Self-contained materials
Distilled water.
Sampler: 5ul, 10ul, 50ul, 100ul, 200, 500ul, 1000ul.
Oscillators and magnetic stirrers, etc.
safety
Avoid direct contact with the stop solution and substrates A, B. Once exposed to these liquids, rinse with water as soon as possible.
Do not eat, drink, smoke or use cosmetics during the experiment.
Do not use your mouth to take any ingredients from the kit.
Operational precautions
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.
The slats not used in the experiment should be immediately put back into the bag and sealed to prevent deterioration.
Other reagents not used should be packaged or covered. Do not mix reagents of different batches. Use before warranty.
Use a disposable tip to avoid cross-contamination. Avoid pipettes with metal parts when drawing stop solution and substrate A and B.
Use a clean plastic container to configure the wash solution. Mix all the ingredients and samples in the kit thoroughly before use.
Wash the enzyme plate should be fully patted dry, do not put the absorbent paper directly into the enzyme standard reaction well to absorb water.
Substrate A should be volatilized to avoid opening the lid for extended periods of 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.
The order of addition of reagents should be consistent to ensure that all wells are incubated for the same time.
The incubation was carried out according to the time indicated in the instructions, the amount of addition and the order.
Sample collection, processing and storage methods
Serum ---- Avoid any cell irritation during the procedure. Use tubes without pyrogens and endotoxins. After collecting the blood, the serum and red blood cells were quickly and carefully separated by centrifugation at 1000 x g for 10 minutes.
Plasma-----EDTA, citrate, heparin plasma can be used for detection. The pellet was removed by centrifugation at 1000 x g for 30 minutes.
The cell supernatant - 1000 x g was centrifuged for 10 minutes to remove particles and polymer.
Storage ------If the sample is not used immediately, it should be stored in small portions at -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. The mouse nicotinamide adenine dinucleotide phosphate ELISA kit should not be thawed by heating at 37 ° C or higher. It should be thawed at room temperature and ensure that the sample is fully thawed evenly.
Reagent preparation
Standard: The serial dilution of the standard should be prepared during the experiment and cannot be stored. Mix the standard shakes before dilution.
Dilution of wash buffer (50 x): 50-fold dilution of distilled water.
Steps
Mix all reagents thoroughly before use. Do not allow the liquid to generate a large amount of foam, so as to avoid adding a large amount of air bubbles during the loading, resulting in errors in the loading.
The number of slats required is determined by the number of samples to be tested plus the number of standards. Multiple holes are recommended for each standard and blank hole. Each sample is determined according to its own quantity, and it is possible to use the double hole as much as possible to make a double hole.
50 ul of the diluted standard was added to the reaction well, and 50 ul of the sample to be tested was added to the reaction well. Immediately add 50 ul of biotinylated antibody. Cover the membrane, mix gently by shaking, and incubate for 1 hour at 37 °C.
Remove the liquid from the well, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 3 times. If washing with a washer, the number of washings is increased once.
80 ul of affinity streptavidin-HRP was added to each well, and the mixture was gently shaken and incubated at 37 ° C for 30 minutes.
Remove the liquid from the well, fill each well with the washing solution, shake for 30 seconds, remove the washing solution, and pat dry with absorbent paper. Repeat this operation 3 times. If washing with a washer, the number of washings is increased once.
Add 50 μl of each of the substrates A and B to each well, mix gently by shaking, and incubate at 37 ° C for 10 minutes. Avoid lighting.
Remove the microplate and quickly add 50 ul of stop solution. Immediately after adding the stop solution, the results should be determined.
The OD value of each well was measured at a wavelength of 450 nm.
Limit
The results above the No. 6 standard are non-linear and no accurate results can be obtained from this standard curve.
Kit performance
1. Sensitivity: The minimum detection concentration is less than the No. 1 standard. The linearity of the dilution. The linear regression coefficient of the sample and the expected concentration correlation coefficient R value was 0.990.
2. Specificity: Does not react with other human cytokines.
3. Repeatability: The coefficient of variation between the plate and the plate is less than 10%.
Result judgment and analysis
1. Instrument value: read the OD value of each hole on a microplate reader with a wavelength of 450 nm.
2. The absorbance OD value is the ordinate (Y), and the corresponding GC1INH standard concentration is the abscissa (X), and the corresponding curve is obtained. The GC1INH content of the sample can be converted into the corresponding concentration according to the OD value from the standard curve.
3. Detection range: 0-80ng/ml
4. Sensitivity: 0.1 ng/ml
The performance of the Baby Monitor Security Camera
The performance of the baby monitor is reflected by its built-in parameters and function combination. The following shows some properties that a high-quality and applicable baby monitor should have. The requirements for baby monitors in this part apply to all audio/video, digital/analog home baby monitors and wireless baby monitors, and are not changed due to the combination mode of the monitoring end and the control end.
The size shall be as small as possible: the small baby monitor (monitoring end and control end) is more convenient to install and carry in the family.Low power display: the baby monitor, especially the monitoring terminal, must work continuously for a long time. Once the product is low in power, it cannot remind parents to charge it in time. When the power continues to be consumed until it is exhausted, the control terminal will be unable to continue to receive any signal.
Voice trigger prompt: Voice trigger means that once the monitoring terminal detects the noise caused by the abnormal behavior of infants or children, it will be displayed at the control terminal through the noise prompt lamp.
The transmission signal frequency can be adjusted: the monitoring end of the baby monitor provides audio and video digital signals to the control end by transmitting wireless signals. If the transmission frequency of the monitoring end is not selected properly, it will produce electromagnetic interference with other household appliances, digital equipment, etc., which will affect the normal use of the product. Therefore, it is necessary to select a baby monitor that allows users to adjust the frequency of the transmitted signal at the monitoring end.
Adjustable volume: a friendly baby monitor will provide a volume selection knob or button at the control end. When parents are in an environment with loud background sound, they can hear the activity sound of babies and children by turning up the volume; When it is late at night or in a quiet environment, the low volume will not affect the normal rest of the people around you when you are answering normally.
VOX function: VOX is the abbreviation of sound activation start. The monitor with VOX function is in standby mode under normal conditions, and the screen and horn stop working, thus reducing the energy consumption of the product; Once it is detected that the monitored object has an abnormal sound exceeding the set standard, it will immediately flash its alarm lamp at the control terminal or directly light up the screen, and simultaneously play the audio and video information sent by the monitoring terminal.
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Monitoring end extension: The baby monitor with a friendly user experience should allow users to use one control end to manage the activities of several monitoring ends at the same time, especially in some families with multiple children. This good scalability not only reduces the trouble of carrying multiple control ends at the same time, but also reduces the objective additional expenses for these families.
Temperature display: some products will have the function of realizing the current indoor temperature, and even can set a critical value. Once the room temperature exceeds or falls below the critical value, an alarm signal will be sent. This is a very considerate function for temperature sensitive newborn children or sick children.
Intercom function: The high-end baby monitor allows parents to have a direct conversation with the baby (child) at one end of the monitoring end through the control end, so that children who are alone can provide psychological comfort. If children have any temporary needs or words, they can be known without bothering the guardian to run to the child.
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Power supply mode: The baby monitor with built-in battery can automatically switch to the battery power supply mode when the external AC power supply is suddenly stopped, so that the user will not be affected to continue to monitor the child in the power failure environment.
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