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Description
Rat GSH-PX 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 specimens using EDTA or heparin as anticoagulants and centrifuge them 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℃, but repeated freezing and thawing should be avoided. 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 1 mL of universal diluent to the lyophilized standard, let stand for 15 minutes to completely dissolve, then gently mix (concentration is 800 mIU/mL). Then dilute to the following concentrations: 800 mIU/mL, 400 mIU/mL, 200 mIU/mL, 100 mIU/mL, 50 mIU/mL, 25 mIU/mL, 12.5 mIU/mL, and 0 mIU/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 800 mIU/mL standard working solution into the first EP tube and mix thoroughly to make a 400 mIU/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 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 Gpx1 Glutathione peroxidase 1 (GSH-PX) 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 Gpx1 Glutathione peroxidase 1 (GSH-PX) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Gpx1Glutathione peroxidase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glutathione peroxidase (GSH-Px or GPx) is a family of enzymes with peroxidase activity. Their primary biological role is to protect the body from oxidative damage. The biochemical function of glutathione peroxidase is to reduce lipid hydroperoxides to their corresponding alcohols and free hydrogen peroxide to water. Several isoenzymes are encoded by different genes, differing in their cellular location and substrate specificity. | |||||||||||||||||||||||||||||||||
| 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 | 12.5-800 mIU/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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★★★★★ 5
Durable Toy for Active Dogs
This ball is strong enough to handle daily play while keeping dogs entertained. It provides stimulation that helps reduce boredom and restlessness. The material feels tough but safe for dogs, and the interactive design makes it more exciting than a regular ball. It’s also lightweight enough for indoor or outdoor use. A smart choice for pet owners looking to keep dogs occupied.
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Reviewed in the United States on November 13, 2025
★★★★★ 5
Bright, easy to clean toy
This ball is a fun little addition for my big dog. The bright blue color really helps, it is easy to spot under the sofa or behind furniture. The outer shell feels durable, not soft rubber but springy enough to handle his chewing without marks. When switched on, it rolls around by itself and makes my dog chase it all over the living room, which is a good way to burn his energy on rainy days. The internal switch is hidden, which I like because my dog cannot open it with his teeth. It is simple to turn on and off, and so far, the movement works as described. The texture also resists hair sticking, and I can rinse it under water, and it looks clean again in seconds. For the price, it serves its purpose and gives my dog a short burst of excitement each day.
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Reviewed in the United States on October 10, 2025
★★★★★ 5
Fun and Tough Interactive Toy for My Big Dog
This interactive dog ball has been a great addition for my large dog. The ball’s material is very durable and holds up well even with heavy chewing. My dog usually destroys toys within days, but this one has lasted weeks and still looks great. The automatic play modes keep him active and entertained, especially when I’m busy. He loves chasing it both indoors and outdoors, and it helps release his energy. The battery life is decent, and the operation is simple. Overall, it’s a fun, sturdy, and smart toy that really keeps my dog busy and happy
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Reviewed in the United States on October 11, 2025
★★★★★ 5
Finally, a Toy That Keeps My Dog Entertained and Active
I bought this interactive dog ball for my German Shepherd, who is very energetic and can be quite aggressive with his chewing. I've tried countless toys that either failed to hold his attention or broke within a week, but this ball is incredibly durable. The durable E-TPU material feels tough and flexible, yet lightweight, making it easy for him to carry around. The bright blue color makes it easily recognizable indoors or out, and I can spot it at a glance.
The ball bounces and rolls in unpredictable patterns, keeping him alert and focused. When he tires of chasing, I switch it to slow mode so he can push the ball around easily. I've noticed he's calmer after playing in the evenings.
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Reviewed in the United States on September 27, 2025
★★★★★ 5
Great way to keep my dog entertained!
My German Shepherd gets bored easily and tends to chew through most toys, but this interactive ball has been a lifesaver. The durable E-TPU material really holds up against his strong jaws. I love that it has 3 different play modes – it keeps him curious and busy for hours, both inside the house and in the backyard. A perfect mix of durability and fun!
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Reviewed in the United States on September 26, 2025