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Description
Mouse 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: 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 the specimen using EDTA or heparin as an anticoagulant and centrifuge 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. Cell Supernatant: 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 and mince the tissue. 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 the 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. Preparation before testing: 1. Please remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of gradient standard working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, and then gently mix (concentration is 10ng/mL). Then dilute to the following concentrations: 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.3125ng/mL, 0.15625ng/mL, and 0ng/mL. Serial dilution method: Take 7 EP tubes and add 500uL of universal diluent to each tube. 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 step to mix thoroughly. 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 IV-C capture antibody. After incubation and washing, the sample is developed with the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the IV-C content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse IV-C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Collagenase is a protease, an endopeptidase, that specifically recognizes the Pro-X-Gly-Pro sequence (a sequence that occurs frequently in collagen and is rarely found in other proteins) and cleaves the peptide bond between the neutral amino acid (X) and glycine (Gly). While many proteases can hydrolyze single-chain, denatured collagen peptides, collagenase is the only one that can degrade native triple-stranded collagen fibrils, which are widely present in connective tissue. Collagenase is divided into four types: collagenase I, II, III, and IV. Type IV collagenase is used to isolate pancreatic tissue and islets. | |||||||||||||||||||||||||||||||||
| 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 supernatant, tissue homogenate, etc. |
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4.4 ★★★★★
Based on 1957 reviews
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Product Reviews
★★★★★ 5
Muy buen producto
Color: Black, Size: 25-Feet
Muy buen producto 100% recomendado y a tiempo
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 14, 2025
★★★★★ 5
Direct Connection
Color: Black, Size: 25-Feet
Received the package in good condition. This wire is a decent quality. I made a set of jumper cables with it and they work excellent.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 24, 2026
★★★★★ 5
bought twice to make custom UPS setup battery leads, not disappointed
Color: Black, Size: 25-Feet
bought this item(25ft 2awg) twice, the first time I did was to create some quick connect cables between my batteries and my distribution block to my 4kw invertor(24v 165a peak calculated load)
I may have oversized due to knowing that the bigger you go the less you stress components and loose power to voltage drop
these cables are a nice high-grade rubber jacket and the 30awg strands are ideal for maximum flexibility while remaining fairly easy to terminate
this cable also includes a backer paper wrap that allows for easy stripping(of the rubber) without too much strand loss damage(from cutting too close to the core)
overall, would buy again, like I did, if I need larger runs(say for feeding an RV solar AC distribution panel) I would buy this brand again, in the 100ft range or so
THHN is not ideal for flex connections(strands are way too stiff) but a couple runs of welding cable like this inside something like "smurf tube" would be a perfect high-power temporary connection system, or just a way to isolate a large battery stack used for a solar setup
in fact I may do that someday, set up a pallet of LiFePO4 batteries with a large quick connect, and just recharge the massive array(s) at a secondary location where solar is more ideal(or other renewables)
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 5, 2024
★★★★★ 5
Custom wires for winch solenoid pack relocation.
Color: Black, Size: 25-Feet
Wire is nice. Flexible. Seems good. However the inner liner is not moisture blocking as I assumed from the pictures, it's paper.. still seems like a good price per foot.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 14, 2020
★★★★★ 4
Good, flexible (considering it's 2 GA) wire
Color: Black, Size: 20-Feet
Good, flexible (considering it's 2 GA) wire. Lugs soldered on well (using solder pot + flux). The only reason I didn't rate it 5 stars is because it's expensive. Made in USA was a +
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 22, 2017
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