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Description
Rat EPHX1 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 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 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 Epoxide Hydrolase 1 (EPHX1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Epoxide Hydrolase 1 (EPHX1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Epoxide hydrolase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Epoxide hydrolase 1 (EPHX1) is an enzyme encoded by the EPHX1 gene. It is primarily present in the endoplasmic reticulum membrane of nucleated cells. In mammals, its expression is generally highest in the liver, followed by the adrenal glands, lungs, kidneys, and intestines. It mediates the sodium-dependent transport of bile acids into hepatocytes. Androstene oxide and oxiranetriols have been shown to be endogenous EPHX1 substrates. It also metabolizes the endogenous plant protein 2-arachidonoylglycerol to arachidonic acid, potentially playing an important role in endogenous plant protein signaling pathways. | |||||||||||||||||||||||||||||||||
| 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 | 31.2-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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Product Reviews
★★★★★ 4
Great Pillow! Could use some improvements
We really like this pillow! It is a good firmness, and helps fill the gap that is often between our headboard and mattress.
Only complain is that almost all of the buttons have fallen off. We have had this for about 2 months.
You also can’t take the cover off of it to clean, which would be a nice feature.
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Reviewed in the United States on February 1, 2026
★★★★★ 5
Pretty, but very practical.
Size: Full, Color: Mint Green
The color was exactly what I expected from the photo. It arrived in an unbelievably small package, but expanded quickly once I opened the bag. I waited the recommended 24 hours to be sure it was done. I am pleased that the length is a perfect fit, and I like that it comes up to the back of my neck when I'm sitting in bed. It is a bit softer than I originally imagined, but not so soft that I feel like I'm sinking through it. It's very pleasantly supportive. The fabric is like a soft ribbed corduroy with piping along the outer seam. It looks nice. Both ends have a pocket that is barely noticeable, but is the perfect size to keep your phone close by. I'm temporarily homebound and spending a lot of time sitting in bed, so I'm very glad I invested in this pillow for comfort and convenience. If you ever have trouble with pillows working their way under the edge of the headboard, and into the dusty space between the headboard and the wall, consider this problem solved.
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Reviewed in the United States on January 23, 2026
★★★★★ 5
Measure your bed and space, I did and ordered the perfect fit!
Size: Queen, Color: Navy
Love this wedge pillow. I bought it for our boat stateroom bed. It fits perfectly as beds on a boat are always odd sizes. It is very comfortable allows me to sit up in bed to read. Also keeps our bed pillows from sliding back. Good quality, color and size.
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Reviewed in the United States on May 12, 2026
★★★★★ 4
Comfy, Supportive, and Great for Lounging in Bed
This wedge-style headboard pillow has made sitting up in bed much more comfortable. The shape offers full upper-back support, with just the right amount of firmness to keep you propped upright without feeling stiff. It’s especially helpful for reading or watching shows in bed, and it stays in place well against a flat headboard.
The purple color is rich and vibrant in person, and the removable cover is soft to the touch with a smooth finish. We really appreciated that the cover zips off easily and can be machine washed—it held its shape and color after laundering. The pillow also has a nice weight to it, so it doesn’t feel flimsy or slide down easily.
This is a great addition for anyone who likes spending time sitting up in bed, whether for reading, working, or relaxing. It also doubles nicely as a decorative accent when the bed is made. The quality feels solid, and the size works well across the width of a queen bed without overwhelming the space. Id say it is ok for the price. It does retain warmth though
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Reviewed in the United States on August 1, 2025
★★★★★ 5
Thick, Supportive, and Cozy for Reading or Nursing
Size: Full, Color: Olive Green
This wedge pillow has been a great fit on my daughter’s bed. It’s super soft and cozy, and at the same time offers great support for sitting up in bed. The pillow is nice and thick, so you don’t sink into it — perfect for reading, studying, or lounging without needing multiple regular pillows.
I tried it out myself and was surprised at how well it also works as support while nursing an infant. It props you up at just the right angle and feels stable without shifting around.
The removable, washable cover is a big plus, and the olive green color is subtle yet stylish. I’m impressed enough that I’ll either be purchasing one for myself soon or adding it to my Christmas list.
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Reviewed in the United States on September 22, 2025