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Description
Rat ANGPTL2 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. 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. 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. 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 utilizes 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 angiopoietin-like protein 2 (ANGPTL2) 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 angiopoietin-like protein 2 (ANGPTL2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Angiopoietin Like Protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Angiopoietin-like protein 2 (ANGPTL2), also known as angiopoietin-related protein 2 or ARP2, is a protein encoded by the ANGPTL2 gene. It maintains tissue homeostasis by promoting adaptive inflammation and subsequent tissue remodeling. Excessive ANGPTL2 activation caused by chronic stress promotes the breakdown of tissue homeostasis caused by chronic inflammation and contributes to the development of metabolic diseases. It also plays a role in angiogenesis, tissue repair, obesity, atherosclerotic disease, and ultimately carcinogenesis. It is a secreted glycoprotein with homology to angiopoietin and can exert its effects on endothelial cells through autocrine or paracrine actions. | |||||||||||||||||||||||||||||||||
| 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 homogenates and other biological fluids |
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4.7 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 4
Medium size preferred.
Size: Large
I purchased both the large 4" and the medium 3" balls. My 80 lbs German shepherd much prefers the medium size. Show loves to play fetch with it and its lasted well!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 10, 2026
★★★★★ 5
Wash first
Color: 25 Pack (Value-Toys)
These are excellent and very well made for a 1 and 1/2 lb Chihuahua girl. I don't know how much they would hold up for a bigger dog but they're perfect for a little one. We always wash all of her toys first and I didn't notice that there were garbage bags in there. Which ended up being a good thing because they were scented. Washing help to get most of the scent off and it took the scent off of the toys. She hates the scent. Me too. It was nice to have the doggy bags though and we would prefer them not scented. We didn't dry the toys and we let them air dry which is a good thing cuz it might have ruin the bags. The bags are a little small and short compared to the normal bags that you buy but overall this is a really good value and she loves all of the teethers and ropes very much. Very well made. Nothing has fallen apart and she has chewed it a lot. I don't know how it would be for a bigger dog but for one and a half pound puppy Chihuahua it's perfect
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Reviewed in the United States on February 13, 2026
★★★★★ 5
My little Chiwoxy (Chihuahua & wire terrier) loves them all!
Color: 25 Pack (Value-Toys)
I was a little nervous to order this set of toys, I worried that they would be sort of junky or dollar store quality. They definitely are not!
My little Mia Bella loves this toy set. Her favorite thing to do is for me to stuff them all in the box they came in, so she can rummage through and each of them out one by one to play with. Empty box, starts all over again. There is a nice variety of soft plush squeakers, soft crinkly toys, ropes with rubber shapes on the ends, ropes with knots, and different kind of balls (some rubber, some rope, some soft plastic spikes). My puppy is very small and I was worried they would be too big for her - they are the perfect size and she loves each and every one of them. Also included were some rolls of poo bags and a plastic holder to hook onto your leash to hold them - I was happy these were included, I hadn’t noticed them on the description, so they were a surprise.
I feel like these toys are a great value and provide a small dog with a lot of fun things to play with. The variety is great so you’re not having to spend money on buying a whole bunch of different things. I am very happy with this purchase, and I highly recommend it to anyone else with a smaller sized dog. I think they might be too small for a very large dog but who knows, they are likely going to be fun for any dog to play with.
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Reviewed in the United States on November 14, 2023
★★★★★ 5
Great selection of dog toys at a good price!
Color: 25 Pack (Value-Toys)
Lots of great toys for your little little dog at a reasonable price. Even comes with doggie poop bags. At any of my local dog shops just one of the ropes would cost me the same as the whole box of toys.
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Reviewed in the United States on June 9, 2026
★★★★★ 4
Excellent Variety Pack - Dogs Love These Toys
Color: 25 Pack (Value-Toys)
This 25-pack is fantastic value with excellent variety—ropes, squeakers, and different textures keep dogs entertained. They're colorful, well-made, and my dogs love them. The main downside is the chew resistance on the rubber toys isn't the best; aggressive chewers can break through them fairly quickly. That said, for the price and the sheer number of toys you get, it's still a solid buy. Perfect for rotation and keeping dogs mentally stimulated. Just know that heavy chewers might go through the rubber ones faster than expected.
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Reviewed in the United States on May 14, 2026
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