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Description
Mouse AACase ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 an acetyl-CoA carboxylase (AACase) 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 acetyl-CoA carboxylase (AACase) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Acetyl-CoA carboxylase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Acetyl-CoA carboxylase (AACase) is a biotin-dependent enzyme that catalyzes the irreversible carboxylation of acetyl-CoA to produce propionyl-CoA through its two catalytic activities, biotin carboxylase (BC) and carboxyltransferase (CT). AACase is a multi-subunit enzyme in the chloroplasts of most prokaryotes and most plants and algae, and a large, multidomain enzyme in the cytoplasm of most eukaryotes. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 8 reviews
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Product Reviews
★★★★★ 3
Spray stops working
Size: 5 Ounce (Pack of 2)
I’ve been buying this product for awhile and keep experiencing issues. It protects your skin well but is sometimes hard to spray and half the time the nozzle stops working while there is still product in the bottle.
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Reviewed in the United States on November 27, 2024
★★★★★ 5
Perfect
Size: 5 Ounce (Pack of 2)
Goes on easy, protects well from the sun.
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Reviewed in the United States on September 10, 2024
★★★★★ 5
This has become my go to daily sunscreen
Size: 1.7 Ounce (Pack of 1)
After first purchasing a year ago, this has become my daily go-to facial sunscreen while others just sit taking up space in my medicine cabinet.
I have sensitive skin and severe reactions to chemical sunscreen and have been searching for the perfect mineral sunscreen for several years after starting tretinoin made my reaction to chemical sunscreen intolerable. I only recently figured out I am also highly sensitive to chemical boosters found in a lot of mineral sunscreens. Especially Butyloctyl salicylate. That one is a no-go for me and until I learned about chemical boosters found in a lot of "100 % mineral" sunscreens, could not figure out why some of my sunscreens made my face flush red and feel like it was on fire when I was in the sun. Lab Muffin is my hero.
My skin is dry, mature, and sensitive with some lines/wrinkes, large pores, and rosacea. At first I thought this Sun Bum tinted mineral would be too drying as it has a very light texture vs being creamy. But it is not and the texture makes for a light but effective formula.
PROS:
- Mineral formula with no chemical boosters.
- Does not settle in pores and wrinkles. After applying it settles slightly in lines, but a quick blend with a blending brush and it is good for the day.
- Blends easily into hairline and brows.
- Does not sting my eyes when I sweat and seems relatively sweat proof.
-Does not transfer tint to clothes once it dries down. Even when I wipe away sweat.... nothing seems to come off. It is the only tinted mineral I have tried with little to no tint transfer.
- Blends easily with a very soft silky texture that allows you to blend without any tugging on the skin like a lot of mineral formulas.
- Matte finish but with a subtle glow.
- Tint provides light coverage to even out skin, though does not cover everything. As someone who rarely wears makeup I appreciate the subtle coverage which does not look overdone but helps my skin look better.
- Does not emphasize occasional tretinoin dry patches. Some of my other sunscreens (even tinted) seem to make dry patches look worse. This formula covers them without emphasizing (or irritating) them.
- Does not feel heavy on my skin. I can't stand feeling sunscreen on my face (or anything which is the main reason I do not wear much makeup) and this formula is so light once it is on I forget it is there.
-Light coverage lasts all day if I don't sweat. Needs some reapplication when sweating.
-Does seem to provide SPF 30 coverage. With reports about sunscreens underperforming lately, this one does appear to work. I can burn in less than 15-20 minutes in full sun, but no problems with this sunscreen when reapplied after sweating or long sun exposure (I work outdoors as a field biologist, so this one is huge. While I usually wear a hat, it does not cover everything as I cannot use a wide brim hat for my work.)
CONS:
- The tube. It is not awful, but I always seem to get a build up of product in the cap no matter what I do. This seems like a waste of product, but is really a minor complaint.
- It does separate, but states clearly to shake before use. It seems to mix easily, so this is not really a con, just a warning to give it a good shake.
- Would love an option for an SPF 50, but again, minor complaint.
Overall, this is by far the best mineral facial sunscreen I have found so far for my particular skin and is used daily. The only reason I ever use anything else is to try to use them up. Other brands I have tried include: Undefined R&R sun serum SPF 50 ( a little too "glowy", emphasizes pores, and has niacinimide which I have to use with caution), Naked Sundays Collagen Glow mineral sunscreen SPF 50 (heavier and feel it on skin, but is my go-to when I must have SPF 50), CoTZ Flawless Complexion SPF 50 (feels heavy and not waterproof. Also even though I can'y find any chemical boosters in ingredients, something in this one makes my skin feel like it is on fire when in the sun), Cocokind daily SPF 32 (ok for a non tinted formula as not much whitecast, but does feel heavier than I would like), Murad City Skin Age Defense SPF 50 ( just too expensive), Koh Gen Do Watery Spa UV SPF 50 (my go-to non tinted formula these days though I still feel it on my skin and not waterproof and think it has been reformulated recently), Vanicream Facial Moisturizer SPF 30 (too thick and heavy), Hero Force Shield SPF 30 (was my favorite next to the Sun Bum until I realized the reaction in the sun was from the chemical booster Butyloctyl salicylate 😩), Agency SilkScreen SPF 50 (was ok, a bit too greasy and not much coverage, but went to back of pack as it has chemical boosters), Haruharu Wonder Black Rice Pure Mineral SPF 50 (I loved this formula and purchased several times before finally learning the reaction in the sun was from chemical boosters), Cetaphil Sunscreen Face Drops SPF 50 (quit using due to flaming face in sun as this one also has chemical boosters), and Etude Sunprise SPF 50 (only used ocassionally due to fragrance, but liked this formula until I clued into chemical boosters).
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Reviewed in the United States on November 15, 2025
★★★★★ 5
Love love love!
Size: 1.7 Ounce (Pack of 1), Size: 1.7 Ounce (Pack of 1)
I’m a big hyram fan and after I found out dear clairs and other Korean chemical sunscreen might not be as effective as claimed, I bought this product off of his recommendation. I was a little nervous about a few things.
For one I have the worlds Oiliest skin. I felt like dear klairs (my old sunscreen) did leave a bit of shine but it had controlled my oil better than any other sunscreen I tried... until now. This is the closest my skin has ever looked to “normal” as far as oiliness goes so props to that!
I also was hesitant on this product because of the tint but also didn’t want a white cast. I would say I’m light to medium with an olive undertone and I think it matches my skin really well! It almost looks like I have makeup on bc my skin tone much more even , but it doesn’t cover breakouts. I happened to have a few pimples (already had them ,not caused by the sunscreen) so you can see that the sunscreen doesn’t cover them completely but blurs/evens out the clear part of my skin. There is a slight white cast but I think the tint offsets it. . I couldn’t speak for other skin tones but I think if you’re light to medium/olive it matches really really well.
Finally I love the texture, it’s almost velvety and doesn’t pull on the skin which I hate in a face sunscreen. Not too thick not too watery, and just feels nice to apply. So I don’t really have anything bad to say about this product, but I’ve only had it two days so I’ll have to see if it causes any breakouts ect. But so far I would highly recommend!
Update: I have been using this for well over a year now, and I can never go back to anything else! It doesn't cause breakouts even when I've slept with it on by accident. I use it instead of a primer under my foundation, and it works amazing. I literally have no complaints; I have not gotten a sunburn since I started using this. I just reapply every few hours when I'm at the beach/pool. I love reapplying because it controls my oil and gives that blurred bb cream effect, I'm also able to put it under my eyes/on my eyelids without burning my eyes. I was never someone who got compliments on my skin, but that changed when I started using this along with the rest of my skincare routine (Shoutout Hyram!) I'm so glad I found this product!
2024 Update!
Still loving and using this sunscreen! I decided to try a couple of different brands earlier this year just because I hadn't ever branched out after finding this product, but this remains the best-tinted sunscreen for oil control and pore blurring I've ever tried!
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Reviewed in the United States on March 20, 2021
★★★★★ 5
Great tinted physical sunscreen
Size: 1.7 Ounce (Pack of 1)
This is my new favorite sunscreen. It goes on matte, but is still easy to put on/not pasty. The tint is enough to get rid of the ghost look but not so much that it looks like fake tanner. It's a bit greasier feeling than I'd like but it doesn't look greasy at all so I'll take it.
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Reviewed in the United States on March 31, 2026
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