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Description
Human 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: 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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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| Sensitivity | 0.05 ng/mL | |||||||||||||||||||||||||||||||||
| 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 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 | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Collagen Type I Alpha 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Type I collagen alpha 1 (COL1A1), also known as alpha-1 collagen, is a protein encoded by the COL1A1 gene. It is a fibrillar collagen found in most connective tissues, including cartilage. Collagen is a protein that strengthens and supports many tissues in the body, including cartilage, bone, tendons, skin, and the white part of the eye (sclera). The COL1A1 gene produces a component of type I collagen, called the proviso α1(I) chain. This chain combines with another proviso α1(I) chain and a proviso α2(I) chain (produced by the COL1A2 gene) to form the type I procollagen molecule. These three rope-like procollagen molecules must be processed by enzymes outside the cell. Once processed, they arrange themselves into long, thin fibers that cross-link with each other in the space surrounding the cell. The result of these cross-links is the formation of very strong, mature type I collagen fibers. The functions of collagen include rigidity and elasticity. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.6 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
I FINALLY found the perfect flavored water!
Flavor Name: Best Sellers Variety Pack, Size: 16 Fl Oz (Pack of 12)
I initially ran across this water b/c it was on sale at my local grocery store and it was NOT what I was expecting at all! I genuinely dislike drinking plain water regardless if it's from the tap, filtered in any way or bottled, regardless of the brand. I will only drink regular water if I absolutely have no other choice.
Over many years I have tried numerous types of flavored waters (regular or carbonated), flavored electrolyte drinks, flavor packets or dissolving tablets, etc. If it's out there I've probably tried it. I am also very attached to sugar, so I didn't mind if they were heavy on the sugar/flavor profile, but I was still looking for something that was more on the pure water side as it's healthier, but also something that would quench my thirst and hydrate me instead of leaving me feeling like I needed more.
I will, at times, order water w/ a lemon wedge and do have some packets that give a very light flavor of lemon or orange in comparison to everything else I listed above. For some reason these were "sufficient", but still not what I was really looking for.
HINT says it all in its name. It gives you a perfect amount of infused flavor rather than syrups, powders, artificial flavoring, etc. It will definitely throw you off for a second the first time you try it b/c it truly isn't like anything else out there that I've tried and is delightfully unexpected. The first time I tried it I THOUGHT it was going to be a disappointment b/c initially I thought it tasted like regular bottled water. Where is the flavor here? But in a matter of a second I tasted the infused flavor profile and it was wonderful. I went around my house and had others try it, knowing they were expecting the same thing I did, and they too were impressed and surprised.
I am still amazed at the way this water tastes and I now order by the case. Because this water is so perfectly balanced and refreshing I have almost completely stopped drinking anything else. For the past 6 weeks or so I have continued to only drink HINT (admittedly I have purchased 2-20oz bottles of soda as a treat), but I do not have any desire to drink anything else. I actually drink water now, and I can't believe I'm saying that.
If you are a water drinker that wants a little bit more, I suggest this product. If you are like me and have spent decades looking for water that is flavored, but still hydrates and refreshes like regular water, look no longer. HINT is unique and bottled perfection.
For people who are conscious of the planet or your body is a temple, HINT also holds a gold standard. The bottles are BPA-free, making the bottle and cap 100% recyclable. Ingredients are only purified water and natural infused flavors. And there are a lot of flavors to choose from. HINT is vegan and free of sugar, sweeteners, MSG, nuts, soy, gluten and preservatives.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 6, 2024
★★★★★ 5
⭐ ⭐ ⭐ ⭐ ⭐ Refreshing flavored water with zero sugar and clean ingredients
Flavor Name: Best Sellers Variety Pack, Size: 16 Fl Oz (Pack of 12)
⭐ ⭐ ⭐ ⭐ ⭐ Refreshing flavored water with zero sugar and clean ingredients
I decided to try the Hint Infused Bottled Water Best Sellers Variety Pack 16 fl oz Pack of 12 because I was looking for a healthier alternative to sodas and sugary drinks. After finishing most of the pack, I can honestly say it has become one of my favorite everyday beverages.
The flavors are very light and refreshing, which I personally prefer. Instead of tasting like artificial sweeteners or syrup, the water has a subtle natural fruit essence. It feels clean and crisp, making it perfect for staying hydrated throughout the day without extra sugar or calories.
I also like that it contains no artificial sweeteners, zero sugar, and zero calories, which makes it a great option for people trying to reduce sugar intake or follow a healthier lifestyle. The variety pack is nice because it allows you to try several flavors and see which ones you like most.
The 16 oz bottles are convenient to grab on the go, pack for work, or keep in the fridge for a quick refreshing drink.
Pros
• Very refreshing and light taste
• Zero sugar and zero calories
• No artificial sweeteners
• Good variety of flavors
• Convenient bottle size
Cons
• Flavor is very subtle (some people may prefer stronger taste)
Overall, this is a great option if you want flavored water without sugar or artificial ingredients. I will definitely keep buying this variety pack to stay hydrated in a healthier way.
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Reviewed in the United States on March 6, 2026
★★★★★ 5
Best bottled water.
Flavor Name: Best Sellers Variety Pack, Size: 16 Fl Oz (Pack of 12)
This water tastes great and has no sugar or sugar substitutes. Good flavors. Highly recommend. A real treat.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
HINT WATER
Flavor Name: Best Sellers Variety Pack, Size: 16 Fl Oz (Pack of 12)
THIS WATER HAS JUST A SMALL AMOUNT OF FLAVOR. IT DOES TAKE A LITTLE GETTING USED TO BUT AFTER A BOTTLE YOU WILL LIKE IT. IF YOU DONT DRINK MUCH WATER TRY THIS YOU MIGHT LIKE IT OVER TAP WATER.
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Reviewed in the United States on May 2, 2026
★★★★★ 4
Excellent if you like flavored water but not seltzer
Flavor Name: Best Sellers Variety Pack, Size: 16 Fl Oz (Pack of 12)
I don't like seltzer but I LOVE this stuff. Minus one star because I think it's pricey. I could drink a bunch of these each day, but I limit myself to one.
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Reviewed in the United States on January 21, 2026