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Description
Rat COL1a1 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. 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 a capture antibody against Collagen Type I Alpha 1 (COL1a1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Collagen Type I Alpha 1 (COL1a1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Collagen Type I Alpha 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Type I collagen alpha 1, also known as alpha-1 collagen, is a protein encoded by the COL1A1 gene. COL1A1 encodes the primary component of type I collagen, a fibrous collagen found in most connective tissues, including cartilage. Collagen is a protein that strengthens and supports many tissues in the body. The COL1A1 gene produces a component of type I collagen called the tropono-α1(I) chain. This chain combines with another tropono-α1(I) chain and a tropono-α2(I) chain to form a molecule of type I procollagen. These three rope-like procollagen molecules must be processed by enzymes outside the cell. Once processed, they arrange themselves into long, thin fibrils that cross-link with each other in the space surrounding the cell. The result is the formation of very strong, mature type I collagen fibers. | |||||||||||||||||||||||||||||||||
| 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 homogenates and other biological fluids |
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4.0 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
they are gorgeous and functional
Color: Ivory
we got these with our cooktop and it is great value and super easy to clean goes in my dishwasher and love how it cooks and cleans they are perfect and gorgeous in white we been using them on the electric stove and they still look brand new. and they cook everything perfectly
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 8, 2026
★★★★★ 5
A great cooking set for just about any meal.
Size: 11-Piece Set
This is a great set of cookware at a very reasonable price. I was looking to make the switch from using non-stick coating to something more natural to minimize the risk of potential toxins. This stainless steel cook set comes with everything that you need to cook just about any meal. As with any stainless cook set, it does require a different approach to cooking, but it's a good tradeoff to know that you're using steel instead of non-stick coating. The quality feels very sturdy, and I feel like these will last me a long time. They are very easy to clean if you soak and use something like bar keeper's friend for anything that is stuck.
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Reviewed in the United States on April 19, 2026
★★★★★ 5
Everyday use
Size: 11-Piece Set
I've used these pans for about a year now and they are still holding up great. For everyday abuse they do not faulter. Plus they clean easy. These pans are a great way to get name brand quality without breaking the bank.
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Reviewed in the United States on May 10, 2026
★★★★★ 5
Solid design, long lasting, healthy surface for cooking
Size: 11-Piece Set
I bought this exact set in 2021. I am buying more. With the number of reports about the dangers of pans made with non stick coating I decided i would only use cast iron or stainless steel going forward. These are “amazon brand” but I can say after years of use and abuse they are standing the test of time. Multiple moves, toddlers, rough use and they are all fully functional and work great. I LOVE the teardrop handles. It makes it so easy to hold the handle and the hanging holes on the handles are large allowing you to hang it more places on more things you may already have set up in your kitchen. I think anyone who is looking to get away from cheap, chemically pans off the rack should look at this set!! I have 4 years of proof they are not junk but a great long lasting items that seem to be well made. Higher price than buying a $10 pan at walmart but you get so many pots and pans with lids that will last a lifetime. You can also use stainless steel to cook in the oven so more functionality than most pans and pots you buy of the rack. You will just need to learn how to cook with stainless steel pans (check YouTube lots of instruction there).
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Reviewed in the United States on April 27, 2025
★★★★★ 4
Very pretty cookware
Size: 11-Piece Set
This is nice quality cookware for a great price. I have seen cooking shows that tell how to cook in stainless steel without it sticking. Unfortunately, my food still sticks. If I use a lot of oil, it isn’t so bad. One cooking show host says if you get it real hot before you add the oil, it won’t stick. It sticks. I am trying to get away from using Teflon because they say we have a lot of Teflon in our bodies from our cookware. I still go back to my Teflon a lot. But the cookware is very pretty, a nice weight, and cleans up fairly easily. I do soak the skillet before I clean it. I don’t have to do a lot of scrubbing. The sauce pans are great.
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Reviewed in the United States on June 10, 2025
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