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Description
Rat cPLA2 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. 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 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 a cytosolic phospholipase A2 (cPLA2) 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 cytosolic phospholipase A2 (cPLA2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cytosolic Phospholipase A2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytosolic phospholipase A2, also known as cPLA2, is an enzyme encoded by the PLA2G4A gene. This gene encodes a member of the cytosolic phospholipase A2 group IV. The enzyme catalyzes the hydrolysis of membrane phospholipids to release arachidonic acid, which is then metabolized into eicosanoids. The enzyme is activated by increased intracellular Ca2+ levels and phosphorylation, leading to its translocation from the cytoplasm and nucleus to perinuclear membrane vesicles. It has been shown to interact with HTATIP. Mutations in this gene are associated with multifocal stenosing ulcers of the small intestine. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 44 reviews
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Product Reviews
★★★★★ 5
Spicy racks
Item Package Quantity: 2, Item Package Quantity: 2
I bought these as I needed to organize my spice cabinet. I was using a lazy Suzanne and the spices kept falling off them. These fit perfectly in the cabinet,I attached them using the command Velcro stickers that come off easy. I am in an apartment. They are strong enough to allow the shelves to slide out.
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Reviewed in the United States on May 20, 2026
★★★★★ 5
Great product
Item Package Quantity: 2
Fit perfectly in my cabinet and gives me the room and accessibility I was looking for. Being 5'4 and needing a step stool to reach my spices above the microwave was pretty annoying. They are solid, quality racks. Very easy to assemble and adheres well to the cabinet.
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Reviewed in the United States on May 14, 2026
★★★★★ 4
Great Quality, but Check Cabinet Height Carefully
Item Package Quantity: 2
I am so, so sad these iSpecle Pull Out Spice Racks did not fit in my kitchen cabinets. I thought I had checked, but the most my cabinets can take with the nonadjustable shelves is 9", and these are a little over 9.5" tall at the metal rack side. They may not fit where I intended to use them, but they appear to be good quality and really help with organization. They look nice and slide in and out well, so I fully intend to find another way to use them. Maybe under the bathroom sink where smaller items also like to cluster.
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Reviewed in the United States on May 19, 2026
★★★★★ 5
How easy they are. And space saving
Item Package Quantity: 2
These are great. Made such a difference in my cupboards. Would rate as a 10
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Reviewed in the United States on May 8, 2026
★★★★★ 5
Finally solved my spice cabinet chaos
Item Package Quantity: 2
I didn’t realize how much time I was wasting digging through my spice cabinet until I installed these. My spices used to be stacked behind one another, which meant every time I needed a specific seasoning, I had to move half the cabinet around just to find it. These racks completely solved that problem.
What immediately appealed to me was the no-drill installation. I rent my home, so I’m always hesitant to make permanent changes to cabinets. The peel-and-stick setup was surprisingly easy, and I had both racks installed in just a few minutes. Once attached, they felt much more secure than I expected. I was initially skeptical about adhesive mounting, but they’ve stayed firmly in place.
The pull-out design is where these really shine. Instead of reaching into a dark cabinet and searching through rows of jars, I can simply slide the rack out and instantly see everything. It’s one of those organizational upgrades that makes everyday cooking noticeably easier.
I also appreciate the adjustable height feature. My spice collection includes a mix of different bottle sizes, and I was able to configure the racks to accommodate both smaller and larger containers without any issues. The flexibility makes them much more useful than fixed-size organizers.
Another thing I noticed is how much cabinet space they helped free up. By using vertical space more efficiently, my cabinet feels less cluttered and much more organized. I can now find exactly what I need without knocking over other jars or creating a mess.
The overall construction feels sturdy, and the sliding action remains smooth even when the racks are fully loaded. This is one of those products that seems basic but ends up making a bigger difference than expected. If your spice cabinet is crowded, disorganized, or constantly frustrating to use, these racks are an easy solution. They’ve made cooking more convenient, improved organization, and helped maximize storage space. For me, that’s an easy five-star product.
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Reviewed in the United States on May 31, 2026