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Description
Mouse SESN2 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Pre-Assay 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 Sestrin 2 (SESN2) 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 Sestrin 2 (SESN2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Sestrin 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sestrin/2 (SESN2), also known as Hi95, is a protein encoded by the SESN2 gene. This gene encodes a member of the estrin family that is related to PA26. The encoded protein may play a role in regulating cell growth and survival. It may be involved in cellular responses to various stress conditions. Sestrins constitute an evolutionarily conserved family of stress-induced proteins that inhibit oxidative stress and regulate adenosine monophosphate-dependent protein kinase (AMPK)-mTOR signaling. These activities make sestrins important regulators of metabolic homeostasis. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | tissue homogenate |
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4.0 ★★★★★
Based on 606 reviews
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Product Reviews
★★★★★ 4
My doggo's go-to toy!
It's not the most durable of toys (she's on her third in five years) but I will buy her as many as she needs because she is OBSESSED with it!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 5, 2024
★★★★★ 5
Nice
Size: Small, Color: Bunny, Size: Small, Color: Bunny
Cute, dog loves it
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 5, 2026
★★★★★ 1
Defective toy and inconsistent quality.
Size: X-Large, Color: Raccoon, Size: X-Large, Color: Raccoon
My initial review was 5 stars based on my experience with a previous order (blue raccoon) and the fact my puppy was immediately interested in it. However, I did not give it to her to play with until this past Friday (May 21, 2026) because she had the first one I bought. In a matter of 2 days the seam on the back and at the base of the head unraveled and all of the stuffing was scattered all over the room. I thought I’d found some good, high quality, toys for her but apparently this one was defective because the first one had had a lot of use and I’d still in decent shape. This is too much money to pay for a dog toy that only lasts a day or two so be advised they aren’t all good.
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Reviewed in the United States on April 11, 2026
★★★★★ 5
Bigger, louder, crinklier, softer, than expected!
Size: X-Large, Color: Rabbit
My dog was gifted a collection of used toys and he really likes a very long stuffed pig which, unfortunately, another dog that is a destructive chewer, totally destroyed. I decided to break down and buy him another long stuffed toy today and since he's always blissfully watching rabbits in the yard and neighborhood I chose the rabbit version. This thing is huge which is great for him- he's 62 lbs and 7 months old and only half grown. He loves it, it has multiple LOUD squeakers, lots of crinkly spots, and fun ears/legs to grab hold off. He's a big fan and it's made with K9 Tuff Guard which LASTS. Super soft and easy to wash.
Update after 2 months- easy to wash and looks new after a washing. very durable so far, standing up to indoor and outdoor use. He is not a destructive chewer but whips them around, tramples on them, steps on them while running top speed and chomps on the annoying squeaker for hours. one squeaker has a hole and its tune is now slightly muted (thank god)
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Reviewed in the United States on August 24, 2019
★★★★★ 3
Puppy loves! Supposed to be part of a set though??
Style: Coconut Tree
Bought this simply because we named our puppy Coco and was on a roll ordering her toys… she ended up absolutely LOVING this tiny palm tree!! Maybe because of the size and she was able to really handle it, she takes it everywhere with her and loves the squeaker. HOWEVER, the item is a lot smaller than I expected when ordering for almost $10 and the plastic packaging it came in says “not intended for individual sale, part of a pack” so I would not purchase from this seller again. I will maybe try to find this included with the intended set of toys.
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Reviewed in the United States on May 28, 2022