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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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4.5 ★★★★★
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Product Reviews
★★★★★ 3
3 Star Read,
Format: Kindle
This book wasn't bad, but wasn't my cup of tea. It's highly disappointing because the storyline is so original. There is no real conflict or resolution, so the entire thing feels flat.
As a lover of omegaverse books, I know there is a ton of variety out there, and ov is really up to the author. But this one is weird. Omegas have multiple scent glands all over their bodies and go into week long heats every month. Alphas have knots in the middle of their shaft instead of the base, and the knot doesn't always swell, no explanation of when or why. It doesn't engage at all when the mouth is in play.
I also didn't enjoy the author's writing style. Each paragraph is only 1 or 2 sentences long, and the entire book reads very stacato. The conversations are stiff and unnatural feeling. Everything is very repetitive, both in word choice and in thought. The same thing is repeated 3 or 4 times over a single page, multiple times over. I ended up doing so much skimming.
The first 50% of the book is all slow burn, and the last 50% is almost straight mediocre spice. This wouldn't have been all bad if the grammar and spelling errors didn't start at the exact same time. Tongue is repeatedly misspelled in the middle of the spice.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 14, 2024
★★★★★ 5
Cozy Omegaverse
Format: Kindle
This is the true definition of Cozy Omegaverse. LA wedding coordinator meets her pack at the location for a couple’s destination wedding. Low angst because they are scent matched high heat with heats and knots. Everything you love about this genre with very little anxiety. Simply a fun experience to read and a book I will comeback to when I’m in a slump. I simply love reading this book
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Reviewed in the United States on March 19, 2026
★★★★★ 4
Cute Omegaverse story
Format: Kindle
3.75 stars and 2 for spice. Adorable Omegaverse story about a wedding planner who has to go to a small town to plan a wedding for a movie star. While there, she meets her pack. This covers her struggles of not living there but trying to keep her relationship going with her 3 Alphas.
It's a quick read and cute, so I recommend it for people who like light-hearted RH stories.
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Reviewed in the United States on May 31, 2025
★★★★★ 3
real good
Format: Kindle
This was a solid read for the ABO universe. The characters are great. The smut scenes were well written. And the town is so cute I want to go there. It didn’t have much character development it just felt like I was getting to the characters as they were getting to know each other.
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Reviewed in the United States on April 27, 2025
★★★★★ 5
So sweet
Format: Kindle
It’s so unexpectedly sweet that it sneaks up on you. Each character is so unique and charming. I love the openness and honesty among their pack. It really blows me away how wonderful this omegaverse trope plays out.
I received a free copy of this book and am voluntarily leaving a review.
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Reviewed in the United States on March 21, 2026