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Description
Human Anxa1 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 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. 5. 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 an Annexin A1 (Anxa1) 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 level of Annexin A1 (Anxa1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Annexin A1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Annexin-A1 (Anxa1), also known as lipocortin I, is a protein encoded by the human ANXA1 gene. Annexin-A1 belongs to the annexin family of Ca2+-dependent phospholipid-binding proteins with a molecular weight of approximately 35,000 to 40,000 daltons. It is preferentially localized to the cellular surface of the plasma membrane. Its apparent molecular mass is 40 kDa and it inhibits phospholipase A2. Anxa1 has important opposing properties in innate and adaptive immune responses: it suppresses innate immune cells and promotes T cell activation. T cell activation leads to the release of Anxa1 and the expression of its receptor. Under pathological conditions, elevated Anxa1 expression may increase the intensity of TCR signaling through the mitogen-activated protein kinase pathway, leading to a hyperactivated state of T cells. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
Based on 537 reviews
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Product Reviews
★★★★★ 5
Excellent pillow.
Size: Upgrade Queen (2 Pack)
Wow these pillows are fantastic!!! First off they are good quality and sized as described. I'm forced to sleep on my back since I use a cpap every night. They suggest taking some filling out for "back sleepers". But in my case, I am disabled, overweight, and the way I position myself on a pillow is probably unique. I have a little of my shoulders, head and neck on the pillow. I didn't have to remove any filling. It was pretty good at first and after about a few day/nights, everything settled in just perfectly. It's truly one the finest pillows I've owned to date. (Part of my disability is a very painful neck & muscle malady, my head leans far to the right. Basically feels like a crick in your neck that never goes away. It's called tortacollis or dystonia depending who you talk to.) Because of this, I have truly spent hundreds and hundreds of dollars on different types and styles of pillows. A very select few have worked out fairly well over time until I found this one. As I said in the beginning, this is an excellent pillow and a great value as well. Hopefully this will work as well or even better for others. I purchased a 2 pack this time, 1 for home and the other for our RV. Definitely buying. another 2 pack to keep for future replacements. Pillows simply don't last forever. They get funky in time. No matter how clean you try to stay with bedding and personal hygiene, it just happens eventually. Sweet dreams. :~)
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 23, 2025
★★★★★ 1
👎🏽 a
Size: Upgrade Queen (2 Pack)
Incredibly thick and firm pillows. Not great for stomach or side sleepers. They alway end up on the floor cause we find them uncomfortable. Don’t recommend, practically unusable
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 5, 2025
★★★★★ 5
It works! Quality pillow.
Size: Standard (1 Count), Color: Original White (Cooling)
This does actually provide a cooling effect, and it's very comfortable! The accompanying literature is helpful too. I believe this to be a quality pillow for the price.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
Most comfortable pillow I’ve ever slept on
Size: Queen (1 Count), Color: Crescent White(cooling)
This is the most comfortable pillow I have ever slept on. It doesn't mash down like other pillows. Cradles my neck perfectly plus it’s cool. I broke my neck a couple of years ago and this pillow feels very good and comfortable. Definitely recommend to everyone. Ive used it since May and it’s retained its shape.
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Reviewed in the United States on June 6, 2026
★★★★★ 4
Smell
Size: Queen (2 Count), Color: Original White (Cooling)
The fact that there are not more comments on the smell of these pillows has me worried about some of yall. Listen, I LOVE the weight, and the firmness, it’s perfect for that. However, these pillows are RANK. They have a very strong mildew-ish smell that lingers. I took the outer cover off and washed it and let the pillow air out for a few days, the smell is still very strong when you press your face against it, which one does because it is in fact designed for that purpose. It smells so bad. It’s nauseating. I can smell it through two pillowcases and blanket that I threw on top of them hoping an extra layer would trap the smell. I’ll bake them in the Texas sun tomorrow and hope that helps, because otherwise these are the perfect pillows for someone who wants to feel like they are hugging a torso but can’t stand the sound/feel of someone else breathing. I don’t know…4 stars, if the smell goes away I’ll come back.
Update: many many days later. Either they don’t really smell anymore or I got use to it. They’ve held their firmness and shape well. I like them.
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Reviewed in the United States on March 9, 2026
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