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Description
Human sCD74 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. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. 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 soluble CD74 (sCD74) capture antibody. 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 soluble CD74 (sCD74) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human soluble CD74 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CD74 (cluster of differentiation 74), also known as the HLA-DR antigen-associated invariant chain or HLA class II histocompatibility antigen gamma chain, is a protein encoded by the CD74 gene. The invariant chain is a polypeptide that plays a key role in antigen presentation. It participates in the formation and trafficking of MHC class II peptide complexes to generate CD4+ T cell responses. The cell surface form of the invariant chain is known as CD74. CD74 is a cell surface receptor for the cytokine macrophage migration inhibitory factor (MIF). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 5
100% need!
It feels so good!! Absolutely love it!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 23, 2026
★★★★★ 4
Versatile spray that works well for sensitive skin
Lately I've been hearing so much about hypochlorus acid and its multiple benefits and uses. Being that I really like Coola products, when I saw that Coola had made a hypochlorous acid face spray, I was eager to give it a try. I'm glad I did!
As someone with sensitive skin that gets irritated from perspiration when I work out, I was hoping this spray might calm my post-workout skin...and it did. The mist comes in an attractive spray bottle. The spray that comes out is at the intensity of somewhere between a fine mist and a coarse spray; it sprays a good amount to wet your face rather than just a very fine coating. Upon spraying on my skin that was red and warm from my workout, I immediately felt a cooling effect. I wiped the spray in to my skin a bit, and it both cleared away the perspiration, and felt soothing to my post-workout irritation. It felt very different from other toners or astringents I'd used, and my face felt clean, refreshed and unexpectedly soft after use.
I later tried the recovery mist on a small area of rash I developed on my neck. I had a similar experience in that it cooled the skin, and my rash appeared minimized, as well.
My only issue with this product is the scent. The spray has a scent that smells like a faint version of chlorine bleach...which was disconcerting, since I'm spraying it on my face! I actually looked it up and apparently that's just the scent that hypochlorous acid has...so it's not specific to just Coola's product. Fortunately, the bleachy scent dissipated quickly.
Overall, I'm happy with this product, and found it useful for my irritated skin. While I tried it for post workout skin and a rash, apparently it can be used for bites, scrapes and acne as well, so it seems to be very versatile. So if you can get over the initial bleachy scent, it's definitely worth giving a try. Priced at $20 for the 3.4 ounce bottle, which should last a while, I find it to offer a good value for the money, as well.
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Reviewed in the United States on March 6, 2026
★★★★★ 5
Surprisingly Effective for Redness & Post-Sun Skin Recovery
This stuff actually works. I’ve been using it after sun exposure, and it genuinely seems to help calm, soothe, and hydrate my skin.
I usually spray it on before applying my hydrating serums and moisturizer, so it’s part of a larger skincare routine, but I definitely feel like this mist plays an important role. My skin feels less irritated, looks less red, and just overall more refreshed afterward.
The spray itself is lightweight and easy to use, with no sticky residue or heavy feeling on the skin. It layers well with other skincare products too, which I appreciate.
Overall, this has become a really nice recovery step in my routine—especially after being outdoors or whenever my skin feels stressed and dehydrated. Sometimes skincare products make very dramatic promises and deliver absolutely nothing but scented disappointment. This one quietly does its job.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Truly refreshing right away
I am someone who gets a red face from even the most slight physical exertion, even when I'm in peak physical shape (I've been this way my entire life), so anything that reduces redness and heat is something I'll pretty much instantly try. I already use several other Coola products (their hair sunscreen is amazing) so this was interesting right away. After going on a brisk walk pushing the stroller (while pregnant by the way), I tried this spray on my face and chest. It was genuinely refreshing immediately! It didn't bother my skin, was a good amount of product in one spray, and didn't have any kind of weird smell. Honestly for $20, I would buy this repeatedly every spring/summer, especially in the Midwest! I'm considering ordering an extra bottle just for my daughter to take with her to summer camp. Pro tip: store in the fridge for even more refreshing-ness!
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Reviewed in the United States on April 17, 2026
★★★★★ 4
Amazing ability to calm red, itchy skin but pricier than alternatives
When I saw this product, I was a bit skeptical. I have extremely dry skin that’s prone to red, itchy spots but it seemed unlikely a misty spray would be anywhere near as beneficial as the creams and ointments I’ve turned to over the years. Since I’ve used other COOLA products and really liked them, I decided to give it a try and I was pleasantly surprised. I tried it on a couple of red patches on my face. There’s an immediate feeling of cooling relief (which leads me to believe this would also be a great solution for a burn), but what I found amazing was the redness decreased significantly and immediately. After a few minutes, I was able to put on makeup, forgoing concealer on those spots. I also tried it on a red, itchy spot on my leg, again calming the itch and decreasing the redness.
Just because it seemed unwise to start using hypochlorous acid, the active ingredient in this spray, knowing nothing about it, I did a bit of online research. Apparently, not only is it considered safe and recommended by dermatologists for eczema like conditions, it also is antibacterial and can be used to keep cuts clean and help them heal. I’m sold on the concept of this spray. My one hesitation is that it seems to be more expensive than other similar options – particularly when you do the math to determine cost per ounce. I am a real fan of the COOLA brand, but given that my skin is such that I’ll probably use this regularly, the cost differential may become prohibitive.
I would highly recommend this product for anyone who wants it on hand for the occasional sunburn or blemish. For those like me who would benefit by using it once or more per day, while still singing its praises as to efficacy, I would have to qualify the recommendation due to price.
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Reviewed in the United States on March 4, 2026
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