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Description
Rat THBS1 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000g for 15 minutes at 2 8C within 30 minutes of collection. Remove the supernatant for testing or store at
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 200 ng/mL). Then dilute to the following concentrations: 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 200 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 100 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 utilizes 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 a thrombospondin 1 (THBS1) 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 thrombospondin 1 (THBS1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Thrombospondin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The thrombospondin (TSP) family is a group of structurally related secreted proteins widely distributed in the extracellular matrix of various tissues. It comprises two subfamilies: TSP-1/2 and TSP-3/4. TSP-1/2 has an inhibitory effect on angiogenesis. TSP-1 was first isolated from the platelet membrane by Baenziger et al. in 1971. It is secreted by normal fibroblasts, endothelial cells, smooth muscle cells, brain glial cells, and bladder epithelial cells. It binds to the membrane receptor CD36 and activates Fyn and Src tyrosine kinases, which are involved in endothelial cell proliferation. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200ng/mL | |||||||||||||||||||||||||||||||||
| Applications | plasma |
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★★★★★ 5
Excellent, prompt, zero hassle protection
I purchased a nice 3D printer for my grandson. He was thrilled and he and his dad used it frequently. The printer developed a serious problem but, thankfully, I had also purchased a three-year protection plan with Asurion. We shipped the printer to Asurion via UPS since they readily agreed to repair it or refund the purchase price. After only a few days, Asurion emailed me that the printer was not repairable. They sent me a link that enabled me to send the full purchase price to my Amazon account. I used it to purchase another printer. I am happy and my grandson is thrilled! I should add that the extended protection did not cover sales tax, nor did it cover shipping cost.
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Reviewed in the United States on March 8, 2026
★★★★★ 5
Strongly recommend
I recently used the services of this company, and I was completely satisfied. They work very professionally: whenever I reached out, they quickly resolved any issues. I strongly recommend that anyone who purchases electronic devices register their product with this insurance. It is truly reliable.
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Reviewed in the United States on June 8, 2026
★★★★★ 5
Cheap Insurance - Easy Claims Process!
I bought this warranty plan for a Dell monitor about 3.5 years ago. About a month ago, it started to give me some intermittent problems (very fast flickering & image retention). I tried everything I could to troubleshoot the issue, but it just kept coming back.
I had some issues with logging into my Asurion account because I used an EDU email, which I no longer have access to. I sent a message to Asurion here on Amazon and they sent me my plan details and everything I needed to make a claim within 24 hrs.
Once I was able to log into my account online, the claims process couldn't be faster/easier. I used the chat function and provided the rep with the issues I was having with my monitor. The rep immediately emailed me a pre-paid UPS label to send my monitor to one of their repair facilities. I got another email when my monitor was received stating that repairs could take up to 5 business days. However, the very next day I got another email stating that my monitor couldn't be repaired and they would be reimbursing me the full purchase price. I received another email with an Amazon gift card for the full price of the monitor within minutes.
This is my first experience with Asurion and I'm very impressed. The customer service is top notch. If I ever buy another expensive electronic item from Amazon, I'm definitely buying an extended warranty from Asurion.
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Reviewed in the United States on April 19, 2025
★★★★★ 5
Better pricing than BestBuy’s Geek Squad extended warranty!
Asurion is on of the best extended warranty put there! bar none! I used to buy only from BestBuy because of their Geek Squad extended warranty. But it’s way more expensive compared to Asurion. although with Asurion they require so much paper work and pictures first.
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Reviewed in the United States on December 26, 2025
★★★★★ 4
They refunded the amount. We just had to send it to them 2 times before they did.
We bought a printer and it was having issues from the beginning. We thought at first it had to do with the WIFI, so we didn't say anything, but then more things were going wrong. I talked to the manufacturer, who said it needed to be replaced but we had to go through Asurion. Asurion "fixed" it and sent it back but we had our doubts. Sure enough, it was within the same month we were sending it in again.
This time they just gave us an amazon gift card for the entire amount we had paid. Thankful for the warranty. They ultimately did the right thing, I just wish we could have gotten it done faster.
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Reviewed in the United States on October 9, 2023