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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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4.8 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Another great story from this author!
Format: Kindle
I absolutely loved this book.
Colette’s story was so sad yet sweet in the sense that yes she ran away from her controlling parents after hearing about arrangements being made for her. But sweet because fate was working in her favor because she ran into one of her mates on the bus. Turns out she had two more back in the town she was heading for.
Dylan, Luca, & Hunter turned out to be exactly what Colette needed because the ease with which she felt comfortable around was just perfect.
This was a fast burn with a pretty quick paced story as well, especially at towards the end, it all picked up pretty quickly towards the end.
I absolutely loved the sweet factor & the epilogues were perfect.
Amazing job!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 19, 2025
★★★★★ 4
Cute read
Format: Kindle
This is a shorter book about a fox shifter who is mute but has telepathy for those she establishes a close emotional bond with. She leaves a rather cold home when she turns 18 and takes a train across country, running into one of her mates on a bus. It's a fated mates book, there's instant attraction but it takes a little while to give into the feelings. It's a fairly low spice level, more like a sweet palate cleanser. It's a standalone but there will be more books set in the same town.
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Reviewed in the United States on March 2, 2025
★★★★★ 5
beautiful story
Format: Kindle
As always a treat for your imagination. Beautiful people with even more beautiful souls. This was a great short story and a lovely quick read with a happy ever after. Thank you for writing such a delightful story. If it was a 1000 pages I would have loved it just as much. Great job…
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Reviewed in the United States on March 23, 2025
★★★★★ 5
So much feelings!
Format: Kindle
Collette was not voiceless in the least and to say she is by far my fave FMC of all time right now it's no understatement. Best 1st read of the year ever!! I love how possessive each of her men were and OMG Lena is the best BFF and girl could ask for! I'm pretty sure I need this gorgeous book on my shelf pronto!
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Reviewed in the United States on February 1, 2025
★★★★★ 5
A Fox Amongst the Wolves
Format: Kindle
Collette has been forgotten about her whole life. Her parents don't bother with her, others don't try either once they realize she uses sign language to speak, and she's a shifter surrounded by humans that hate her kind. On her 18th birthday, she runs away. She takes the train and then bus wherever it can get her as far from home as possible. A missed bus brings her and a mysterious but kind man to Willowdale. What happens next is a story of fated mates, shifters, magic, new best friends and found family, and a stolen child. I'm sad this was a shorter read but only because of how much I loved Collette, Dylan, Hunter, Luca, Lena, and the others in their community. I love how detailed the characters are even with the shorter story. You can tell their characteristics easily, and the lead in to book two is done so well. I loved this insta love book with just the right amount of drama.
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Reviewed in the United States on July 10, 2025