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Description
Mouse AGC 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. 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 Aggrecan (AGC) 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 Aggrecan (AGC) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Aggrecan ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aggrecan, also known as cartilage-specific proteoglycan core protein (CSPCP) or chondroitin sulfate proteoglycan 1, is a protein encoded by the ACAN gene. This gene is a member of the clockican (chondroitin sulfate proteoglycan) family. The encoded protein is a component of the extracellular matrix in cartilage tissue, which supports cartilage compression. Aggrecan is a proteoglycan, or a protein modified with large carbohydrates. This protein is 2,316 amino acids long and, due to alternative splicing, can be expressed as multiple isoforms. It is also a high-molecular-weight proteoglycan. It exhibits a bottle-shaped structure in which chondroitin sulfate and keratan sulfate glycosaminoglycan (GAG) chains are attached to an extended protein core. Aggrecan has a molecular mass greater than 2,500 kDa. The core protein (approximately 300 kDa) has approximately 100 GAG chains attached to it. Aggrecan consists of two globular domains at the N-terminus (G1 and G2) and a single globular domain at the C-terminus (G3), separated by a large extended domain (CS) that is heavily modified with GAGs. These two major modifications are themselves arranged into distinct domains: a chondroitin sulfate domain and a keratan sulfate domain. Aggrecan also plays a crucial role in organizing the extracellular space between neurons in the brain. Through interactions with linker proteins and tenascins, aggrecan binds to hyaluronic acid, forming large aggregated complexes on the cell surface. | |||||||||||||||||||||||||||||||||
| 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 | 78.1-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.7 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 4
Great toy but if your dog is an aggressive chewer, please think twice.
Color: Crinkle Chicken (Brown), Size: Large
I received this on May 23rd and on June 8th, after playing with this thing every day, our recently acquired 36 lb dog finally ripped it open and got out all the plastic and the squeaky part. Lily Belle will miss you, chicken.
Lily loooooved this toy! She'd toss it in the air, chew on it, pull it with her teeth etc. but as much as she loved it, we won't replace it. It's just not durable enough for her.
If your dog is an aggressive chewer, you might want to try a different toy. I hate that this lasted just a smidge over two weeks because Lily seemed to truly enjoy this toy.
RIP, squeaky chicken!
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Reviewed in the United States on June 9, 2026
★★★★★ 5
Great Value Toy
Color: Crinkle Duck (Blue), Size: Large, Color: Crinkle Duck (Blue), Size: Large
This toy has been one of my dog’s absolute favorites. He gets excited every time he sees it and will carry it around the house, toss it in the air, and keep himself entertained for a long time. The shape and texture seem perfect for him, and it quickly became one of his go-to toys over everything else in his basket.
What I really like is that it holds up reasonably well for the price. It’s definitely not indestructible, and after a lot of play, it will sometimes start to rip—usually around the feet first since that’s where my dog tends to grab and chew the most. But honestly, considering how affordable it is, I don’t mind replacing it when needed. It’s inexpensive enough that rebuying feels worth it because he genuinely loves it that much.
I’d rather keep buying a toy he is obsessed with than spend more on something that just sits untouched. It’s fun, cute, and keeps him happy, which is what matters most. Overall, I’d definitely recommend it for dogs who love plush toys and playful chewing.
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Reviewed in the United States on April 28, 2026
★★★★★ 5
Best affordable squeaker toy
Color: Crinkle Chicken (Brown), Size: Large, Color: Crinkle Chicken (Brown), Size: Large
my dog is obsessed with this toy, this has became his new comfort toy and after having this for about four months the legs are falling off but it’s very durable. The squeaker also moved from the head of the chicken to the body of the chicken but that’s after it was played with every single day. I love that it has two squeakers and it’s the perfect toy for my dog.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
My dogs favorite toy EVER
Color: Crinkle Chicken (Brown), Size: Large
I have a Belgian Malinois who loves any and every toy, but this one is definitely her favorite. She will walk around the house constantly carrying it in her mouth, which is honestly adorable to watch.
The squeakers only lasted a couple of months, but I don’t really consider that a downside—I’m kind of grateful they didn’t last forever given how much she plays with it. It still remains one of her most-loved toys even without them.
My only word of caution is that if you work from home, this toy makes a lot of crinkling noise. And I mean A LOT. It’s great for keeping the dog entertained, but it can definitely get a little frustrating for humans trying to focus or take calls.
Overall, it’s been a big hit in our house and easily one of her favorite toys.
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Reviewed in the United States on June 8, 2026
★★★★★ 5
Super cute
Color: Bunny (Mauve), Size: Small
Super cute and fuzzy. Definitely perfect size for a puppy and not too loud. The fur is very soft and the nose is sown on not a hard piece that can be chewed on. Very good quality too.
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Reviewed in the United States on June 3, 2026
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