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Description
Rat IV-C 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 specimens using EDTA or heparin as an anticoagulant and 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. 3. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 4. 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 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 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 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 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 IV collagenase (IV-C) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Type IV collagenase (IV-C) 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 IV collagenase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Collagenase is a protease consisting of a single polypeptide chain of approximately 1,000 amino acid residues. Collagenase digests native collagen fibers, commonly found in connective tissue, and is therefore often used for tissue dissociation. Collagenase preparations contain the activities of several proteases, including collagenase, casein, clostridase, and trypsin (Kessler & Yaron). Type IV collagenase, which contains a low level of trypsin activity, is often used for pancreatic islets (Taguchi et al.) and in applications where maintaining receptor integrity is crucial. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatant |
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4.7 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 5
Great workout mat
Color: Green
It’s very thick and soft so it makes exercising a lot easier. It rolls easily ready to store. And it seems very durable. It is also flexible for certain exercises for balance.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 29, 2026
★★★★★ 5
Very sturdy. Great Quality
Color: Navy Blue
This is a very dense, thick mat. It's great if you have bad knees or just need/want a thicker mat. The carrying strap with velcro closures is easy to use.
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Reviewed in the United States on April 21, 2026
★★★★★ 5
Great cushioning, easy storage
Color: Blue
I am so happy I bought this. I do back stretches, and need to be on the floor.. Getting up & down is hard at 77, and carpets are hard. This helps sooo much! I don’t hurt my knees when getting down or getting up. The mat is comfortable and provides plenty of padding. It is easy to roll up and store with the included strap. I am tall, and there is extra length, so my head and feet are on the mat. This was a good purchase! This has not torn, and wipes down easily. It doesn’t attract dog hair, which is another reason I wanted a mat.
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Reviewed in the United States on March 21, 2026
★★★★★ 5
Great Knee Support
Color: Blue
The 1-inch thickness provides exceptional support and cushioning, making it ideal for a variety of exercises beyond just yoga.
The high-density foam used in this mat is a game-changer. It not only cushions my joints but also retains its shape remarkably well.
The carrying strap is a convenient addition, allowing for easy transport wherever I go. Plus, the mat is easy to clean, which is a huge plus after sweaty workout sessions.
While the extra thickness might not suit everyone's preferences, especially if you prefer a thinner mat for certain poses, I find it perfect for my needs. It strikes an excellent balance between comfort and stability, providing the right amount of cushioning without compromising stability during balancing poses.
Overall, the BalanceFrom All Purpose Yoga Mat will become an indispensable part of my fitness routine. Its durability, support, and easy maintenance make it a top choice for anyone seeking a reliable exercise mat for various workouts.
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Reviewed in the United States on November 27, 2023
★★★★★ 5
Perfect For My Routine
Color: Black
I am retired military, so exercise has always been a part of my daily routine. Since the pandemic I have had to adjust, which is hard because the gym has the equipment I have become accustomed to. I didn’t realize how different it was working out there as when I started to work out at home on my hard wood floors (no carpet anywhere). I thought it would be easy but the military exercise mat is entirely too thin and was making my tailbone hurt tremendously, so I had to give up my sit-ups and some other ab exercises. This kind of threw a wrench into my routine and I could start feeling my motivation go away. I decided to take a look on Amazon for a thicker mat. I should have done this long ago because I found this one I’m writing about. It’s the shape of any typical exercise Matt except it’s 1” thick. It appears to be made of a type of foam (not bothering to look at the description right now). When I pulled it out I thought I was going to sink in making sit-ups even harder but it’s firm but not hard. I like it better than what is at the gym. Durability will be hard to determine until I’ve had it a while but I’m definitely excited to start back up on my exercises.
I recommend this mat if you have hard wood floors, bad back, bad knees, etc.
I thought I read some negative reviews about mats this thick because of yoga and balance but I beg to differ. If balancing is part of your exercise routine, then a mat which makes balance harder is exactly what you want. Exercise is supposed to be challenging and you need to work harder to see a difference. If that isn’t your goal or if balance is a medical issue, this may not be the right mat for you. Something you may not think about is I have chronic foot pain and this mat makes it less painful for me to stand or jump.
For me, this mat is perfect.
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Reviewed in the United States on July 26, 2020