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Description
Rat PLA2G2A 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 12 ng/mL). Then dilute to the following concentrations: 12 ng/mL, 6 ng/mL, 3 ng/mL, 1.5 ng/mL, 0.75 ng/mL, 0.375 ng/mL, 0.1875 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 12 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 6 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 a capture antibody against Phospholipase A2, membrane associated (PLA2G2A). 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 Phospholipase A2, membrane associated (PLA2G2A) in the sample. Absorbance (OD) is measured at 450 nm using a microplate reader to calculate sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Phospholipase A2, membrane associated ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Group IIA phospholipase A2 (PLA2G2A), also known as PLA2 or sPLA2, is the enzyme encoded by the PLA2G2A gene. It is a member of the phospholipase A2 family (PLA2), which constitutes a diverse family of enzymes in terms of sequence, function, localization, and divalent cation requirements. The gene product belongs to group II and contains a secreted form of PLA2, an extracellular enzyme with a low molecular weight that requires calcium ions for catalysis. | |||||||||||||||||||||||||||||||||
| 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 | 0.188-12 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 10 reviews
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Product Reviews
★★★★★ 4
Good for gentle dogs, not so much for chewers
Color: Green
My pup loves this and it definitely keeps him entertained, but the fleece ties are not the strongest so if you have a strong chewer be ware that it will not last too long (he's been playing with it for a day and we've lost 3 so far)
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Reviewed in the United States on April 19, 2026
★★★★★ 3
Hard to fill and time consuming but fun concept
Color: Green
Cute and fun concept for my dog. He enjoyed pulling out the strips and getting the treats it was hard to shove the pieces back in after rolling treats into it. I used small training treats and it still made it bulky and hard to shove back in. If the strips were shoved too far in it’s hard to get it out. I was suprised my 120 lb malamute didn’t rip the treat strips so the quality of the fabric holds up well. The ball material itself is thick and quality and he didn’t rip the rubber apart either. The strips once out seemed difficult for him to unroll and he lost interest after pulling a few out and he enjoys his other puzzle toys more.
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Reviewed in the United States on February 8, 2026
★★★★★ 5
A Tail-Wagging Triumph: The Dog Snuffle Ball – Interactive Puzzle Dog Toy
Color: Green
Behold, the Dog Snuffle Ball—a toy so ingenious, so joyous, it might just be the pinnacle of canine entertainment! This isn’t merely a toy; it’s a symphony of stimulation, a carnival of enrichment, and a therapist rolled into one delightful ball of wonder.
At first glance, it appears unassuming—a soft, colorful orb that’s almost too cute to resist hugging yourself. But the moment you tuck treats or kibble within its folds, magic happens. Your dog transforms from a curious onlooker to an intrepid explorer, embarking on an epic quest for buried treasure. It’s as if their primal instincts awaken, and suddenly, they are the Sherlock Holmes of the snuffle world.
The brilliance of the snuffle ball lies in its multifaceted benefits. It’s a mental workout, sniffy fun, and slow feeder all rolled into one glorious package. Training your dog’s sense of smell? Check. Keeping them occupied for hours? Double check. Preventing boredom and destructive behavior? Oh, you bet. Watching your dog experience pure, unbridled joy as they unravel its mysteries? Priceless.
And let’s talk versatility: This marvel isn’t just for dogs. Cats, rabbits, and even the occasional curious human (yes, I’ve tested it myself!) can find delight in its interactive design. Its soft, durable material ensures that pets of all sizes and ages can join in the fun without fear of injury or wear.
The snuffle ball is also a hero of stress relief. For pups prone to anxiety or mischief, it channels nervous energy into productive play, transforming would-be chewers of shoes and shredders of cushions into happy, focused furballs of contentment.
To the masterminds behind the Dog Snuffle Ball, I offer a standing ovation. You’ve created a toy that’s not just fun—it’s transformative. It turns every mealtime or treat session into an adventure, fostering happiness and health in our beloved companions.
The verdict? Every pet needs this toy. Not some, not most—every single one. Buy it, toss it, and prepare for a tail-wagging extravaganza of sniffing, pawing, and pure bliss. 🐾✨
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Reviewed in the United States on January 19, 2025
★★★★★ 5
Great product
Color: Green
It was fun to watch my dog, get the food out of the rolls of fabric
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Reviewed in the United States on May 31, 2026
★★★★★ 5
Great puzzle, even better complimentary squeaky carrot
Color: Red, Color: Red
Our dog is a Jack Russell mix and needs near constant interaction. In the winter, it gets so cold that we can't always get him out for long enough periods of time to satisfy his poor bored little brain.
I've tried so many puzzles. He gets frustrated and bored quickly, and ends up just barking at them.
One game that he loves in winter though, is what we call "the blanket game" and it's when we hide one of his favorite toys in an old blanket, wrap it all up, and then let him loose to find the toy. He can play this game over and over, for over an hour if we have the energy. So, when I saw this ball, I knew I had to try it.
He loves it. He begs me to let him have it at least once a day and it gives him a good 5-10 minutes of play. He'd play it more if I let him have that many treats, haha.
But I had to say that, true to form, his REAL favorite part of the puzzle ball was the complimentary carrot toy that came with it.
He is obsessed with the carrot.
He carries it everywhere. It goes to bed with him. He never loses it, always knows exactly where Carrot is. He even knows the word "carrot" now, after only a week or so of having our dear, beloved Carrot.
I can ask him, "go get Carrot," and he brings it right to me.
It is the only toy he ever brings me now.
The squeaker is long dead, he killed it within the first couple days, but he still chews it relentlessly, listening for the little air puff sounds it makes now.
So anyway, the ball is great. Love it. But the carrot? Is an epic find. I included a picture so you can see how utterly filthy it is from how much he loves it (don't worry, it's going in the washer).
Thank you for this random carrot. It might be the best thing to happen to our weird little family this winter.
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Reviewed in the United States on February 6, 2024