Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 21 - Jul 26
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
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. |
|||||||||||||||||||||||||||||||||
| 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 |
|
|||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.3 ★★★★★
Based on 20 reviews
Sort
Product Reviews
★★★★★ 5
My dog is a toy destroyer
Color: Consuela the Cactus - Large, Size: Large Dog, Color: Consuela the Cactus - Large, Size: Large Dog
I’ve had a hard time trying to find toys that my dog, Zara loves because she’s actually very picky. Ive gotten plenty of Kong toys that are well known to be indestructible but she wants nothing to do with them. Then I got her ropes and that seemed to be good for a while until she was getting sick in the middle of the night from eating all of the string. Stuffed toys aren’t good because she’ll rip them within seconds and eat all of the stuffing. I felt like I’ll never score the perfect toy for her.
I found the three layered cactus toy and decided to give it a go. It’s clearly designed to be ripped and destroyed to get the ball in the middle which is actually really cute and fun to see all of the layers.
The cactus arms have stuffing and of course that’s the first thing she went for and ate. I had to watch her and make sure I took the stuffing away before she ate it. And before I knew it, she ate half of the first layer fabric lol once she got through the first layer, all she’s been doing is walking around the house with the toy in her mouth. But now she’s tired and passed out on the couch from all of the fun.
This toy won’t last long with a toy destroyer but it sure does give them something to do and it’s a cute toy. I do recommend!!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 26, 2024
★★★★★ 5
Last a while. Will be buying more toys soon.
Color: Consuela the Cactus - Large, Size: Large Dog
Best dog toy I have ever bought. I have three dogs, two at the time I bought this toy. It took them over a year to break through the first layer. The flower comes off first so be careful your dog doesn't swallow it. It still has the second layer so it still looks like a cactus. No stuffing to clean up when they tear into it, which makes it far less messy than other toys. One of my dogs is obsessed with fetch so I use it like a ball inside. The oval shape makes it hard to ensure when it bounces that it will bounce straight. It's bounced into walls and other stuff that I wasn't intending it to hit. Not much of a squeak so I can let them play with it for hours and I don't get annoyed with the noise. Great toy that lasts a while.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 25, 2025
★★★★★ 5
Nice Value
Color: Consuela the Cactus - Large, Size: Large Dog
Very cute! Its soft and feels like it is well made. It is a heavier toy and a tougher one. Not a plush toy. I will order again for our older dogs. Nice value for sure!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2026
★★★★★ 5
Great toy for our best boys!
Color: Consuela the Cactus - Large, Size: Large Dog
You can never go wrong with BarkBox toys and this was no exception. Our dogs like toys that have squeakers and that they can tear into. This style of toy fits the bill. The design is cute as well. I’ve had this in the house for months and, while it’s definitely seen better days, it’s still intact and they haven’t gotten to the inner layer. (Not for want of trying.)
The perfect type of toy for our fur babies.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 10, 2026
★★★★★ 5
Survives 7 months so far.
Color: Christmas Consuela, Size: Large Dog
It is still intact after nearly 7 months of constant play. My blue heeler normally destroys toys in just a few weeks.
Consuela has exceeded expectations of survival with an extremely chewy, destruction prone dog.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 6, 2026