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Description
Human trypsin ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with trypsin capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of trypsin in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Trypsin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Trypsin is a serine protease from the PA superfamily found in the digestive systems of many vertebrates, where it hydrolyzes proteins. Trypsin is formed in the small intestine and becomes trypsin when its proenzyme form, trypsinogen produced by the pancreas, is activated. It primarily cleaves peptide chains at the carboxyl side of the amino acids lysine or arginine. Trypsin is used in many biotechnological processes. This process is often referred to as trypsinization, and proteins digested/treated with trypsin are said to be trypsinized. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 2081 reviews
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Product Reviews
★★★★★ 1
BOUGHT 2 AND BOTH TERRIBLE
Style: -20-Degree, Size: 50-Foot, Style: -20-Degree, Size: 50-Foot
I bought two of these products 6 months apart from each other (march 2024 and October 2024) both brand new out of the box leaked from this hose connection fitting and is not fixable by replacing the standard hose gasket. Thought maybe it was just the first one was just a dud. It was not, the 2nd one was exactly the same and leaked out of BOTH connection fittings aswell. Yes i tried new gaskets, thread tape / dope, trying to tighten it a bit more, pressure reducer down to abt 35 psi. None of which worked. This product has potential of being good but they used such a low quality connection that it ruins the whole thing. Havent gotten to the point where Ive used it below freezing for more than 1 night so I cant comment on the freeze proofness of it. Tested it with an electrical meter and it uses 690W when plugged in and below the temp rating. I ended up having to repair the hose myself because I couldnt order a chinese brand one instead without running out of time before freezing temps (photographed repair of what is necessary for this to funtion propertly out of the box) but for the price point this hose is at I expect it to last at least 2 years without issues at least, I especially DO NOT expect it to leak being brand new out of the box. If you are going to do the repair you have to carefully cut the insulated sleeve back a few inches without cutting the hose or heated wire and then cut the original hose connector off. At that point you can use pretty much any hose repair threaded connector with a hefty hose clamp nice and tight (dont strip the hose clamp threads though). After spending another 15 min on the repair and $10 in parts you will then have a functional hose for at least 1 week. We'll see how long it lasts now.
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Reviewed in the United States on November 7, 2024
★★★★★ 5
Hasn't froze yet
Style: -20-Degree, Size: 25-Foot
Ive been using this for 2 winters now. Its not lost its color, its not kinked, its still rigid and sturdy. It rolls up compact and fits into storage easy when warm. It can be difficult when cold. Easy to use, you just need typical wall power and a water source. Its been buried in nearly a foot of snow and single digit cold and hasn't froze yet.
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Reviewed in the United States on May 9, 2026
★★★★★ 5
A 5 year sentence
Style: -20-Degree, Size: 50-Foot
I’ve had this hose for 5yrs and it just sprang a leak and now I need to replace it lol. I’d say that’s pretty damn good for a hose that gets constant daily use. When the weather gets into the sub zeros it does freeze up, even with added heat tape and foam insulation around it all buried about half a foot underground.
All in all, even with the freezing I’d say it was a great hose, very durable, not too stiff but a durable firmness, it connected very easily and with the thread tape on it never leaked.
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Reviewed in the United States on July 8, 2025
★★★★★ 5
Smart, Well-Built RV Sewer Hose Support
Size: 30-Foot
I’m quickly becoming a fan of Camco products—they’re smartly designed, well-constructed, and a great value. This 30-ft Sidewinder sewer hose support performs perfectly, providing a smooth and stable transition from the RV’s discharge hose to the septic system. I especially appreciate the Velcro storage strap that keeps everything neatly together when not in use. It’s flexible, durable, and easy to handle. Highly recommend this product to any RV owner looking for a reliable and practical hose support solution. Tip: Always get a longer support than your actual sewer hose length so that you can double up the support were needed. The support can never be too long, but too short could cause flow issues during your camping trip.
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Reviewed in the United States on January 10, 2026
★★★★★ 5
Light weight and easy to stage!!
Size: 10-Foot
I recently purchased the RV Sewer Hose Support with a collapsible design, and I must say, it has exceeded my expectations in terms of convenience and functionality. Here's my review based on my experience with the product:
Pros:
Collapsible Design: The standout feature of this sewer hose support is its collapsible design. When not in use, it folds down easily and takes up minimal storage space in my RV. This is a significant advantage, especially in small RVs or when you have limited storage capacity.
Stability: I have fully extended the support multiple times, and I'm impressed by its stability. The product doesn't wobble or tip over, even when I have my sewer hose filled with waste. This sturdiness gives me peace of mind, knowing that my hose is well-supported and less likely to leak or cause any mess.
Durable Plastic Construction: The choice of durable plastic for the construction of this sewer hose support is a smart decision. The plastic is tough and can withstand the outdoor elements, including exposure to sunlight, rain, and occasional rough handling. I feel confident that this product will last me a long time, making it a worthwhile investment.
Cons:
Honestly, I haven't come across any significant drawbacks with this product. It has served its purpose exceptionally well, and I have no complaints about its performance.
Overall, I highly recommend the RV Sewer Hose Support with a collapsible design. It addresses all the essential aspects you would want in such a product: ease of storage, stability, and durability. The collapsible feature makes it a breeze to pack up, and the stable support ensures smooth sewage flow without any mishaps. The durable plastic ensures that it can handle the rigors of outdoor use, making it a reliable addition to any RV owner's toolkit. Whether you're a seasoned RVer or just starting, this product will undoubtedly make your RVing experience more convenient and hassle-free.
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Reviewed in the United States on July 26, 2023