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Description
Human ES 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: 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: 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. 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. 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. 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. 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. 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 enzyme conjugate are sequentially added to microwells pre-coated with endostatin (ES) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of endostatin (ES) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Endostatin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Endostatin (ES) is a naturally occurring, 20 kDa C-terminal fragment derived from type XVIII collagen. It acts as an anti-angiogenic agent, similar to angiostatin and thromboelastin. Endostatin is also a broad-spectrum angiogenesis inhibitor, interfering with the pro-angiogenic effects of growth factors such as basic fibroblast growth factor (bFGF/FGF-2) and vascular endothelial growth factor (VEGF). It is produced by proteolytic cleavage of collagen XVIII by proteases (such as feline proteases). Collagen XVIII is a member of a family of polypeptides characterized by interruptions of the triple helix to generate multiple domains. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 1545 reviews
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Product Reviews
★★★★★ 5
Improved my foot health
Size: Medium, Color: Color Mix
I’ve been dealing with athletes foot for decades and using daily maintenance medication for longer than many people reading this have been alive. I generally don’t have any issues other than I always at least one or two nails that are cloudy and brittle. I’ve tried synthetic blends in the past but this just seemed to make things worse so I always go back to cotton.
I was been reading about the benefits of wool so I thought I would try these socks. I had several clouded nails at the time but in just a few weeks of wearing wool socks I noted subtle improvements. At the time of writing this it has been over 4 months of wearing wool socks exclusively and all my nails are completely clear and healthy! I honestly can’t believe it since I haven’t had seen completely healthy nails since I was a teenager. I do continue to use daily antifungal maintenance medication but the only change has been the wool socks.
I think part of the issue was the cotton socks being soaked with sweat when I took them off at the end of the day regardless of the season or activity levels. The wool socks, however, are usually dry. Even after taking them out of a hiking boot on a hot day on the trail they will feel damp but not soaked. Consequently, they dry out completely on a rock while taking a short break. By staying dry they don’t bind up and cause hot spots like cotton socks. That, and I think they just distribute the heat better than cotton and I wouldn’t be surprised if they are actually cooler on hot days. Of course, in cooler temps they are warmer and I did appreciate how much so until the other day when I wore some cotton socks that felt like I was wearing ice cubes on my feet when I was walking around my hard wood floors at the end of the day.
With these particular wool ankle socks I did note a hole around the toes with a couple of the socks after a few washes. My experience with cotton socks is that even the smallest of holes will grow exponentially with each use and so I generally just discard them. At first, I was disappointed thinking that these socks would not last but it does not appear that the holes grow like cotton. In fact, I haven’t thrown one wool sock away. My eyes aren’t great at close distant but it seems like the holes I noticed disappeared. Even if they didn’t it doesn’t matter since they do not currently seem to be a problem. I haven’t had these socks long enough to notice any thinning at wear points like cotton socks but I suspect that since they don’t bind like cotton they are less prone to wear.
I now have a dozen pairs of wool ankle socks and I’ve gotten in the habit of putting them in a separate bin for washing otherwise they end up getting tumble dried with the rest of the family’s close. They have been in drier a number of times and I suspect they look a little more aged as a result. If I wash them separately with other delicate items I just lay them out on the floor after washing since they aren’t noticeably wet after a spin cycle and they completely dry out overnight. At this point I would rather put on a pair of wool socks straight out of the washer then put on dry pair of cotton socks since the results would be the opposite at the end of the day.
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Reviewed in the United States on October 21, 2024
★★★★★ 5
Nice look, good value
Size: Large, Color: Grey Mixed(quarter Socks)
Fit great. Cushion on bottom is exactly what I wanted, top and ankle sections are much thinner as I wanted, so as not to be bulky. Expect they will well for walking and hiking.
They did wash well although did lose some of the softness but did not shrink.
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Reviewed in the United States on March 30, 2026
★★★★★ 3
Nice but issue from manufacuter.
Size: Large, Color: Ivory/Beige/Pink, Size: Large, Color: Ivory/Beige/Pink
I received the 6 pack of socks to give as an Easter present. The socks were great colors, soft, and fit well. Will be a little warm during the hotter days. Unfortunately the yellow pair came with a hole already at the toe area.
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Reviewed in the United States on April 6, 2026
★★★★★ 5
My dog LOVES this tiny tug ball
Size: Medium(6")
A lot smaller than I was expecting.. BUT my dog doesn’t mind at all. She seriously loves this thing. It goes flat fairly quickly, but that’s what happens with all of her soccer balls, no matter the size. She’s a pitbul catahoula mix and has a strong bite. Considering my dogs chewing abilities, I’m fairly impressed by the durability of the ball, though she chewed through all but one of the rope handles in about 2 seconds. All that said, it’s one of her favorite toys(she’s 13 years old)
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Reviewed in the United States on April 21, 2026
★★★★★ 5
10/10
Size: Medium(6")
My dog loves this ball. Durable and the straps seem to be strong too. Would highly recommend
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Reviewed in the United States on May 27, 2026