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Description
Rat ICTP 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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| 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 Type I collagen pyridine cross-linked end peptide (ICTP) capture antibody. 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 concentration of Type I collagen pyridine cross-linked end peptide (ICTP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Type I collagen pyridine cross-linked end peptide ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Type I collagen is primarily found in bone, constituting 90% of the organic composition. It is also found in loose connective tissue, alongside other collagens such as types III, V, and VI. Type I collagen also accounts for the largest proportion in these areas. ICTP is the pyridine cross-linked end peptide of type I collagen, participating in trivalent cross-links and released during the degradation of mature type I collagen. The immunochemically intact form of this end peptide can be found in the blood and appears to be derived from bone resorption and the degradation of loose connective tissue. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.8 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
So good I bought two
Size: 6 tier, Color: Black Brush Silver
These shelves are GREAT. Why? They arrive in a compact well protected package, they go up in minutes, are super tall, and they look fantastic. The price is perfect. The components are not 'real' iron pipes (I've built my own iron pipes shelves before and it's extremely heavy), but some kind of lighter weight metal. That's fine by me! The shelves appear to be real wood (!!). If you buy cheap furniture you know what fake wood looks like. You can tell. Guests can tell. This is probably a pretty cheap wood but who cares--it's REAL, is stained nicely with a distressed look, and overall looks amazing once books and items go on it.
If you are deciding between these and some cheaper similar thing, pay a biiiit more and get something with real wood. I just bought a second for my office and they'll hold my books, art, and speakers.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2021
★★★★★ 5
Perfect. Sturdy. Super easy to assemble.
Size: 6 tier, Color: Black Brush Silver
I don’t rate a lot of items but I’m in love with this one. I just received it today. It was packaged incredibly well, took 15 min or less to put together and looks amazing!!! I’m tempted to order another one for another room in the house. It’s sturdy, well-made and I can’t stress how easy it was to assemble. Great product, highly recommend. (The owners even sent a hand-written note thanking me for ordering - what a nice touch!)
Edit: ordered a second one after living first one so much
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Reviewed in the United States on December 30, 2020
★★★★★ 5
Perfect industrial look bookshelf
Size: 6 tier, Color: Black Brush Silver, Size: 6 tier, Color: Black Brush Silver
Surprised how good the quality was for the price. Metal is real metal and wood seems to be real wood. Pretty much twist on and off each section to put together. No question it is a very sturdy piece of furniture. Bought it over other bookshelves because the reviews said it was solid … and indeed it did turn out to be very solid. I’m very pleased and would recommend. Now I just need some things to fill the shelves.
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Reviewed in the United States on May 18, 2022
★★★★★ 5
Looks great & saves space
Color: Rustic Brown and Black, Color: Rustic Brown and Black
I couldn't be happier with the look and the space it's saving me. It came very well packaged, even with corner guards on the wood shelfs. It was very easy to assemble and attach to the end of my pantry in the kitchen. That space was just going to waste. The ladder shelf fit perfect and looks great. I love the color and texture of the wood grain, it really goes well with the black metal rail. It will be a little wobbly until you get it screwed down to the wall but after that, it's good and sturdy. The instructions were easy enough to read but I still found a couple of parts I didn't use and wondered what they were for? The parts do come numbered but only on the packaging? Not on the part itself which would have been more helpful to some extent. So just keep it all together with it's packaging and you'll have no problems. Very happy with purchase.
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Reviewed in the United States on July 31, 2023
★★★★★ 5
Nice and sleek
Color: Rustic Brown and Black
I’m really happy with the bookshelves overall, I only wish they had taller ones available. It was very quick and easy to put together and they have held up well so far. They look great for the price and I especially like that they have a small footprint when it comes to how much room they take up.
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Reviewed in the United States on May 5, 2024
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