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Description
Mouse GS 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 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. 3. 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. 4. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend them 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, the PBS volume can be reduced 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. 5. Cerebrospinal Fluid 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. 6. 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 a glutamine synthetase (GS) 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 amount of glutamine synthetase (GS) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Glutamine synthetase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glutamine synthetase (GS) is an enzyme that plays a vital role in nitrogen metabolism by catalyzing the condensation of glutamate and ammonia to form glutamine. The glutamine synthetase reaction is glutamate + ATP + NH3 → glutamine + ADP + phosphate. Glutamine synthetase utilizes ammonia generated by nitrate reduction, amino acid degradation, and photorespiration. The amide group of glutamate serves as the nitrogen source for the synthesis of metabolites in the glutamine pathway. Other reactions may occur via GS. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cerebrospinal fluid, cell culture supernatant and other biological fluids |
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4.5 ★★★★★
Based on 1148 reviews
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Product Reviews
★★★★★ 5
Holy Grail Moisturizer for Rosacea
Style: Restorative Protective Cream-3.3 Fl Oz
In seeing so many negative reviews I felt compelled to weigh in as this is my favorite moisturizer. I have very dry, menopausal skin. Additionally, I have rosacea which seems to have been a nice gift from said menopause (you can only laugh, right?). It's very common for those of us in our 50's to have a compromised skin barrier and when an issue like rosacea is present, even more so. Because of all this my skin is very intolerant to most moisturizers and nearly all actives (over the last few years I've had to discontinue my retinols and chemical exfoliators). While ingredients like ALA's, petrolatum, mineral oil, parafin/bees wax, and lanolin are well tolerated and known to be quite soothing, they don't hydrate very well as they are more occlusive than moisturizing. Enter this product. Magically, it does both! It's marketed as a protective cream, which again implies occlusive, but it penetrates like no other. Gone is the tight, dry feeling that persisted even when I had a heavy layer of product on my skin (my previous moisturizer was Eucerin Original Healing Cream -- which still has its uses -- because it was the only thing I could put on my skin outside of squalane oil during a flare).
Now, no topical can cure rosacea. However, the right product can calm the skin, repair the damaged lipid barrier, and improve its appearance. This moisturizer does just that and it's become my holy grail. Since starting this I've also added in the Cicalfate Serum which has only aided in repairing my skin. Another Avene product that works great for me is the XeraCalm Balm which helps with itching. I use all three of these daily and they've really helped fix the secondary issues that come with rosacea.
As for the complaints about it being greasy or too heavy. I can see this not working for people with normal or oily skin (or perhaps only in moderation). But my skin loves the heavier weight and because one of the ways I soothe a flare is to keep a tiny mister bottle on hand and repeatedly spray my face so I have to guard against the drying effects of evaporation. These products are so lipid heavy that I can wet my face all day long without worrying about that, though I do reapply throughout the day.
As for the watery consistency issue, I haven't had that problem but it does caution on the packaging not to expose the product to temperatures below 41 degrees F (5 C). I suspect that if it gets too cold either because of the weather or because of whatever warehouse it sat in during transit it will separate. Maybe open the package and before squeezing some out look to see if it has split and then shake it up/message the tube.
So, if you have skin like mine don't hesitate to try this product. I'm leaving this absurdly long review for my struggling brothers and sisters out there with rosacea in hopes that it provides some relief! Happy moisturizing, internet strangers. ;)
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Reviewed in the United States on July 18, 2025
★★★★★ 4
Possible solution to seperation
Style: Restorative Protective Cream-1.3 Fl Oz
I love this product so much, but, like everyone else the water HEAVILY separated. A lot of water just shot out immediately and as i've used the cream it has felt more and more thick bc of that, shaking the tube or not.
POSSIBLE SOLUTION:
I actually think the company could salvage this problem by upgrading packaging to a Cosmetic JAR and then customers could take a toothpick and stir if separation has occurred by the time of purchase. I hope to try this with my next tube, just toothpaste squeezed everything out into a nice cosmetic jar and stir the water back into the product. This is such a simple solution that could really solve the product waste.
This refreshes my skin, doesn't break me out, and is lovely. I break out so easily, and being middle aged my skin really requires a dense help with moisture. <3
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Reviewed in the United States on April 22, 2026
★★★★★ 5
Ordered in spring/summer and it arrived separated too BUT...
Style: Restorative Protective Cream-1.3 Fl Oz
Ok, So I heard great things about this cream and decided to order. I ordered on May 28th and I live on the Gulf Coast of Texas and it is in the low 90s right now. I was worried mine would arrived separated like many others claimed to have gotten theirs, and sadly it did. Warm weather or not, blue water shot out of the tube the minute I squeezed it. Luckily, I read Mia P's review and submerged my tube in warm water (hot water actually straight out of my instant hot water faucet). I left it for about 5 minutes then shook it vigoroously. It seemed to help but was still a bit wonky so I repeated the process again and success! My tube is totally fine now and comes out in a thick, rich, emollient cream as it is supposed to. I will update my review after I use it more but so far I love the way it feels on the skin. It is very thick but spreads nicely and completely absorbs into the skin and leaves no greasy residue which is a big deal because I have oily skin. I purchased this to help with my Retin A uglies (I actually am using Musely with .05% tretinoin) and am hoping it helps with the redness and peeling I am having. I will update my review once I've used it for a while but wanted to post this review as I saw so many people complaining about the separating and just thought I'd mention that you can fix this issue if you have it (and there is a good likely hood that you will).
Update: I've been using this for a week now and I am absolutely in love with this cream! It is the most hydrating and healing cream I've ever used. Usually, thicker face creams like this would cause issues for me with acne or clogged pores as I do have oily skin that can break out if I use products with occlusive ingredients, but this one does not. In fact, my skin has never looked better! This is a must-have if you are using Retin-A/Tretinoin and going through the red/peely phase with your skin. The separation that occurred when I first received it has not been a problem and the cream is staying thick and free from any strange liquidy issues after I put it in hot water as mentioned in the first part of my review.
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Reviewed in the United States on May 28, 2023
★★★★★ 5
Saved My Skin and Healed My Eczema
Style: Restorative Protective Cream-1.3 Fl Oz, Style: Restorative Protective Cream-1.3 Fl Oz
I was having the worst skin winter of my life. I am hypersensitive and my skin started rejecting an expensive line I had used for many years (EL). My skin wasn’t healing even just using AQ balm on it and occasionally steroids when it was extra bad, and the pronounced eczema on my eyelids and neck for months was making me miserable and gross looking. I started researching brands I hadn’t tried for super sensitive skin and saw recommendations for Avene. We were leaving on a vacation and I was desperate to feel better so I ordered this and the matching serum. I started with only this cream and overnight I saw my skin start to heal. Within a few days it was like the last few months never happened. I carefully added the serum and had no issues. I used only these two products for the month I was gone and had glowing skin the whole time. This is thicker than a normal lotion since it is made to protect your skin but if you let it warm up a few seconds it smoothes easily. This will definitely remain a staple in my skin arsenal and I can’t thank you enough for making an actually gentle but effective product! This picture is two weeks after starting the product, smooth skin and confident to take selfies!
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Reviewed in the United States on March 3, 2026
★★★★★ 5
Don’t let the bad reviews deter you from saving your skin
Style: Restorative Protective Cream-3.3 Fl Oz, Style: Restorative Protective Cream-3.3 Fl Oz
I avoided purchasing this product for a while due to the bad reviews. However, it is still highly recommended by several dermatologist for skin barrier repair. Upon doing my own research, the product can separate if it is stored in cold temperatures (41 degrees or less). This can be fixed people!! It literally says it on the bottle/box and there’s even a video that Amazon posted saying this can happen. It does not affect the product. It’s just the thermal spring water and some oil separating but it can be reincorporated.
So I took the risk and ordered this. I’m a first time purchaser. It was already at my door when I was leaving for the gym at 5:30am so I let the package sit in my car for about 1.5 hours (temp was about 70s). I put the tube in a glass of hot sink water for 10 minutes when I got home. I rolled the tube in my hands for a few minutes to mix everything together. Guess what. It came out like a CREAM. No liquid shot out at me. If you can read, do a little research, and can follow instructions, you can get a working product.
I have sensitive skin. It’s been a struggle healing my skin barrier but it was getting better. I decided to do something dumb and try dermaplaning. My cheeks have been very red and irritated since. I did avoid active products for a few days after dermaplaning but it was still irritated. Just with the first time use of this product, it was nearly instant relief of some of the redness. It’s fragrance less which is great. It is currently on sale so I think the value is good for what it is right now. I only used two pea-sized amount for my whole face so I think it will last awhile.
This can significantly help your skin barrier! Do your own research and read the instructions!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2026