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Description
Rat SOD1 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. Cell lysis solution: Gently wash adherent cells with pre-cooled PBS, then digest with trypsin, and collect the cells after centrifugation at 1000×g for 5 minutes; suspended cells can be directly collected by centrifugation. Wash the collected cells 3 times with pre-cooled PBS, add 150-200uL PBS for every 1×10^6 cells to resuspend (it is recommended to add protease inhibitors to PBS; if the content is very low, the PBS volume can be appropriately reduced) and break the cells by repeated freezing and thawing or ultrasound. Centrifuge the extract at 2-8℃, 1500×g for 10 minutes, and take the supernatant for detection. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes, take the supernatant for detection, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 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 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/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 final calculation of sample concentration should be multiplied by the corresponding dilution factor. It is recommended to run replicates for all 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 superoxide dismutase 1 (SOD1) 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 level of superoxide dismutase 1 (SOD1) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Superoxide Dismutase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Superoxide dismutase 1, also known as superoxide dismutase [Cu-Zn] or hSod1, is an enzyme encoded by the SOD1 gene located on chromosome 21. SOD1 is one of three superoxide dismutases. It has been implicated in apoptosis and familial amyotrophic lateral sclerosis. SOD1 is a 32 kDa homodimer formed into a β-barrel with an intramolecular disulfide bond and a binuclear Cu/Zn site in each subunit. This Cu/Zn site accommodates copper and a zinc ion and is responsible for catalyzing the dismutation of superoxide to hydrogen peroxide and disulfide. The copper chaperone (CCS) of Sod1 facilitates copper insertion and disulfide oxidation. Although SOD1 is synthesized and matures in the cell membrane, the partially expressed, immature form of SOD1 that targets mitochondria must be inserted into the intermembrane space. The mature protein is highly stable, but is unstable in the metal-free and disulfide-reduced forms. This is demonstrated in vitro by increased aggregation of SOD1 following metal ion depletion. In disease models, insoluble SOD1 is poorly metallated. SOD1 binds to copper and zinc ions and is one of three superoxide dismutases responsible for destroying free superoxide radicals in the body. The encoded isoenzyme is a soluble protein located between the cytoplasm and mitochondrial membranes. It functions as a homodimer to convert naturally occurring but harmful superoxide radicals into molecular oxygen and hydrogen peroxide. The hydrogen peroxide can then be broken down by another enzyme, catalase. | |||||||||||||||||||||||||||||||||
| 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 | 78.1-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids |
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4.4 ★★★★★
Based on 10 reviews
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Product Reviews
★★★★★ 5
Nice Value
Color: Consuela the Cactus - Large, Size: Large Dog
Very cute! Its soft and feels like it is well made. It is a heavier toy and a tougher one. Not a plush toy. I will order again for our older dogs. Nice value for sure!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2026
★★★★★ 5
Great toy for our best boys!
Color: Consuela the Cactus - Large, Size: Large Dog
You can never go wrong with BarkBox toys and this was no exception. Our dogs like toys that have squeakers and that they can tear into. This style of toy fits the bill. The design is cute as well. I’ve had this in the house for months and, while it’s definitely seen better days, it’s still intact and they haven’t gotten to the inner layer. (Not for want of trying.)
The perfect type of toy for our fur babies.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 10, 2026
★★★★★ 5
Survives 7 months so far.
Color: Christmas Consuela, Size: Large Dog
It is still intact after nearly 7 months of constant play. My blue heeler normally destroys toys in just a few weeks.
Consuela has exceeded expectations of survival with an extremely chewy, destruction prone dog.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 6, 2026
★★★★★ 4
Short-lived but much enjoyed as a chew toy
Color: Consuela the Cactus - Large, Size: Large Dog
I bought two of these for my "granddogs" in the size for Large dogs. The German Shepherd-mix stole the one intended for the border collie-mix and had both toys halfway destroyed within a day--I'm pretty sure they didn't survive much longer after that. Much enjoyed! Maybe I should've bought the Extra Large size, because the Large barely fit in her mouth and she wanted to play tug-of-war, so there wasn't much for me to grab onto. Then the border collie wanted in on the action (she's older, so she needed help tugging!) but there wasn't enough material for her to grab onto without accidentally sinking her teeth into me instead.
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Reviewed in the United States on December 9, 2025
★★★★★ 5
Best dog toy!
Color: Air Head, Size: Large Dog, Color: Air Head, Size: Large Dog
I got this toy for my dog who loves squeakers not only is it a two in one it actually has three toys inside which I think is amazing!! 10/10 recommend
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2026
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