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Description
Human CTRP4 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. Centrifuge the specimen 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. Tissue homogenization: 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 the tissue and mince it. 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 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against C1q and Tumor Necrosis Factor Related Protein 4 (CTRP4). After incubation and washing, the sample is developed using 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 amount of C1q and Tumor Necrosis Factor Related Protein 4 (CTRP4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human C1q And Tumor Necrosis Factor Related Protein 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | C1q and tumor necrosis factor-related protein 4 are proteins encoded by the C1QTNF4 gene. Cq1TNF4, also known as CTRP4, is a secreted protein and one of at least nine analogs of Acrp30/Adiponectin. Proteins in this family contain a globular C-terminal domain similar to that of C1q and the TNF ligand family. C1qTNF4 is the only family member lacking a collagen-like domain but possessing two C1q domains. Mature human C1qTNF4 shares 92.4% amino acid sequence identity with the corresponding region of mouse C1qTNF4. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Powerful
Color: Black, Color: Black
I love it because it doesn’t use batteries — you just recharge it and that’s it. Plus, it’s very powerful and works really well. Honestly, I didn’t expect it to be this good. Great option and excellent value for the price.
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Reviewed in the United States on May 28, 2026
★★★★★ 4
Good product
Color: Black
Good product I like that it’s rechargeable. The only thing I don’t like is that there’s no power button. You have to press it while in use.
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Reviewed in the United States on June 6, 2026
★★★★★ 5
Amazing quality for the price!
Color: Black, Color: Black
⭐⭐⭐⭐⭐ Perfect for My Morning Coffee!
I absolutely love this milk frother! It’s simple to use, works quickly, and creates a rich, creamy foam that makes my coffee feel like it came from a coffee shop. I was surprised by how well it froths both warm and cold milk in just a few seconds.
The frother is lightweight, comfortable to hold, and easy to clean after use. It’s become a part of my daily coffee routine, and I find myself using it every morning.
If you're looking for an affordable way to elevate your coffee experience at home, this is definitely worth it. Highly recommended🌟⭐️
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Reviewed in the United States on June 2, 2026
★★★★★ 5
Fast Charging & Powerful Mixer Works Perfectly!
Color: White, Color: White
I’m really impressed with this little mixer overall. It comes with a universal Type-C charging port, which is such a huge convenience for daily use. I don’t have to carry extra exclusive charging cables around anymore, it matches my usual charging cords perfectly.
The battery life is surprisingly great too. After a single full charge, it lasts for such a long time on daily use, no need to charge frequently at all.
What I love most is its mixing performance. It has strong power and a high working speed. It blends everything super evenly and smoothly in seconds, no uneven mixing or lumpy spots at all. Definitely a high-power, practical daily gadget that totally exceeds my expectations!
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Amazing product, and even more AMAZING Customer Care Team!
Color: All Black Premium Gift Box, Size: Z1 Motor + Batteries, Color: All Black Premium Gift Box, Size: Z1 Motor + Batteries
I don't normally write reviews, however I can't not mention the ABSOLUTELY AMAZING Customer Care experience I had with these folks.
But first, In review of the product itself, which is the 'Executive Series Black Milk Frother with Black Holster Stand'. This is my first 'frother' purchase, and I must say I was skeptical at best to try it, but I opted to purchase it anyway because of all the great things the company stands for. I figured, how good could it really work?! Well, as a matter of fact, it worked quite wonderfully! Within just a few seconds / pulses of the frother, you have beautifully frothed contents. It is simple to operate, with one hand, and just the simple push of a button. (I imagine I could even make whipped cream without dragging out a stand mixer.) The batteries, (2 AA - not included), were simple to load. And, there is a battery pull ribbon so you can also remove them easier, if you utilize the pull ribbon and thread it between the batteries. This device quickly became part of my weekend coffee routine, and I also purchased a lesser model to bring to work. (This 'Executive' one, has a holster stand, which looks nicer, and seemed to cradle the frother, instead of hooking it into the holder.)
I was so happy with the frother, I was sad that one day it seemed to not be working quite as well. I thought .. hmm, maybe the batteries need to be changed? I changed the batteries, and was still experiencing it not really working. (I would press the button, but it didn't start frothing right away, and then seemed less powerful in a way when it did start. But, it did still froth.) After a second day of experiencing the same thing, I reached out to Zulay. They are gracious enough to have ALL of their contact information printed directly on their box, underneath all of the beautiful information about the company, and where / how it began. It was on a weekend, so I chose to send an email describing what I was experiencing, and asking their advice. Based off most recent attempts at Customer 'Service' with other companies, I didn't really expect to hear back. Not only DID I hear back, but I received an email from a human, not just a system-generated response with canned information. They expressed their concern for my item not working, and assured me they would make it right if given the opportunity. They wanted to confirm my order, to make sure they had the same model in stock. Within hours, they sent me a link to replace my item, and would ship it directly to me, no questions asked. No need to return the inoperable item, no need to do anything but literally click the button and wait for it to arrive.
My replacement arrived, and it was the exact model I purchased. It was brand new, wrapped in package .. not some refurbished item. And, much to my shock and appreciation that almost NO ONE does this anymore, there was a hand written card included in the package thanking me for giving their family run business a shot. Well, I thank YOU for sharing YOUR family with us, and for making me feel a little like a piece of yours!
I would highly recommend this product, and would certainly purchase it, or others, in the future. I would certainly also be comfortable gifting this item.
Enjoy!
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Reviewed in the United States on April 13, 2024