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Description
Human a2-MG 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: 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: 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. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge it at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃. However, repeated freezing and thawing should be avoided. UrineOther 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 250 ng/mL). Then dilute to the following concentrations: 250 ng/mL, 125 ng/mL, 62.5 ng/mL, 31.25 ng/mL, 15.625 ng/mL, 7.8125 ng/mL, 3.90625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 250ng/mL standard working solution into the first EP tube and mix thoroughly to make a 125ng/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 an alpha-2-macroglobulin (a2-MG) 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 alpha-2-macroglobulin (a2-MG) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Alpha-2-Macroglobulin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Alpha-2-macroglobulin (α2M) is a large (720 kDa) plasma protein found in the blood. It is primarily produced by the liver, but is also synthesized locally by macrophages, fibroblasts, and adrenal cortical cells. In humans, it is encoded by the A2M gene. α2-macroglobulin acts as an antiprotease, inactivating a wide range of proteases. It functions as a fibrinolysis inhibitor by inhibiting plasma proteins and calreticulin. It also acts as a coagulation inhibitor by inhibiting thrombin. α2-macroglobulin may act as a carrier protein, as it also binds to many growth factors and cytokines, such as platelet-derived growth factor, basic fibroblast growth factor, TGF-β, insulin, and interleukin-1β. | |||||||||||||||||||||||||||||||||
| 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 | 3.90-250ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, urine, and other biological fluids |
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4.3 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 4
Super fun for the dog, but doesn’t last.
Size: Small, Style: Bone, Product Packaging: Standard Packaging, Size: Small, Style: Bone, Product Packaging: Standard Packaging
I just ordered the 3 rd one. The bad thing is that my dog destroyed it in 3 hours. Luckily she spits out all the small pieces of plastic. The good thing is, that she gets so much joy out of it, it makes her so happy and tired that I am a fan! Well worth 9$!
She is 25lb and I didn’t think she was an aggressive chewer before this toy. If your dog tends to eat all the plastic pieces , don’t get it.
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Reviewed in the United States on April 4, 2026
★★★★★ 3
See updated review; destroyed within 15 minutes
Size: Medium, Style: Bow Tie, Product Packaging: Standard Packaging
I'm bummed. My pup destroyed this within fifteen minutes. I thought because it was made by Kong, it would hold up a little better. I also have their black rubber toys and those are great!
Update: I didn't expect to be contacted by the seller but they reached out after seeing my review and offered a full refund which I received quickly. Truly outstanding customer service!!!
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Reviewed in the United States on December 2, 2025
★★★★★ 5
Durable Chew Toy with a Fun Design
Size: Medium, Style: Bow Tie, Product Packaging: Standard Packaging
The KONG CoreStrength Bow Tie Bone is a playful and sturdy chew toy made to keep dogs busy. Its unique bow tie bone shape makes it easy for dogs to grab and carry, while the reinforced core is built to handle strong chewing. The textured surface not only adds interest but also helps clean teeth and gums during play.
It’s tough enough for moderate to heavy chewers and works well for fetch, solo chewing, or tug games. While no toy is completely indestructible, this one holds up longer than many softer toys and gives dogs plenty of chewing satisfaction. Some very aggressive chewers may eventually wear it down, but most dogs will get a lot of use out of it.
Pros: The reinforced core makes it more durable than average toys, the textured design supports dental health, and the shape adds variety and fun.
Cons: Very heavy chewers may still break it down over time, the toy is fairly heavy for small breeds, and it isn’t designed to bounce or fly like other KONG toys.
A strong and engaging chew toy that combines durability with a fun shape. It’s a smart choice for dogs who love to chew and play while also supporting healthier teeth and gums.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 30, 2025
★★★★★ 5
Best chew toy for GSD’s
Size: Medium, Style: Bone, Product Packaging: Standard Packaging, Size: Medium, Style: Bone, Product Packaging: Standard Packaging
One of my dogs has chewed through most chew toys for aggressive chewers with ease. This toy not only is still in one piece but still her favorite toy. We’ve gotten her many Kong toys and this one has definitely lasted the longest. She’s been chomping on it for about 2 years now and looks mostly whole.
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Reviewed in the United States on April 15, 2026
★★★★★ 5
🐶 Unleash the Fun with the KONG CoreStrength Bone: The Ultimate Chew Toy for Mighty Molars! 🦴
Size: Medium, Style: Bone, Product Packaging: Standard Packaging
Okay, fellow dog parents, gather around! Have you ever wished for a chew toy that could survive the apocalypse? Well, the KONG CoreStrength Bone is here to save the day (and your sanity)!
This isn't just a chew toy; it's a canine's dream turned reality. The multilayered core? It's like the Captain America of dog toys – robust, enduring, and ready for any amount of chew-action your four-legged friend can dish out. Long-lasting chew sessions? Please, that's child's play for this toy.
Now, let's get down to the nitty-gritty – dental health. The textured exterior is like a toothbrush disguised as a toy. It's working overtime, cleaning those teeth and massaging those gums while your pooch is none the wiser, lost in a world of chewy bliss.
And if you've got a dog that chews through toys like they're made of butter, this is the holy grail. Made for the toughest of chewers, this bone is like the superhero of dog toys, ready to withstand hours, days, even weeks of relentless gnawing.
The design? Pure genius. It's not just a toy; it's a mental workout, keeping your dog entertained, engaged, and away from your favorite shoes. Plus, it's from KONG – a name that's like the Rolex of dog toys. Quality? Check. Durability? Double check.
Every dog deserves a toy that's as fun as it is functional, and the KONG CoreStrength Bone ticks all the boxes. Made in the USA with globally sourced materials, it's the kind of toy that makes tails wag and pet parents smile.
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Reviewed in the United States on December 27, 2023