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Description
Rat CAPN2 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 frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 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 calpain 2 (CAPN2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of calpain 2 (CAPN2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Calpain 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Calpain 2 (Calpain2) is a protein encoded by the CAPN2 gene. Calpains are calcium-activated neutral proteases and non-lysosomal, intracellular cysteine proteases. This ubiquitous enzyme consists of heterogeneous forms with different large catalytic subunits associated with a common small regulatory subunit. The gene encoding the large subunit of the ubiquitous enzyme, calpain 2, has multiple heterogeneous transcription start sites in its 5' UTR. It has been shown to interact with Bcl-2. | |||||||||||||||||||||||||||||||||
| 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 homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 23 reviews
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Product Reviews
★★★★★ 5
Westies Fav
Size: Small, Pattern Name: Pack of 1
Best Westie ball ever, my dog loves this ball. Exactly her size! 3" tennis balls are too big & she rips the fur off. this silicone-ish ball has no fur & is 2&1/2 "
We have 2" balls from her electronic ball laucher, but she still loves this glow in the dark best!
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Best dog ball in the world!
Size: Large, Pattern Name: Pack of 1, Size: Large, Pattern Name: Pack of 1
My dog is singularly obsessed with playing ball. He would drop a steak to chase a ball. (He'd return to the steak, but not without the retrieved ball. He's not an idiot.) He makes dangerous leaps, crashes into trees, and runs fast enough to catch thrown balls before they even hit the ground. Since he doesn't care about his own wellbeing, it's my job to protect him as much as I can. And a lot of balls are hard enough to cause him pain or dental damage when he mistimes a catch. Enter the ChuckIt Glow Ball!
The minute Cowboy got his first Glow Ball, he no longer had any interest in the dozens of tennis balls scattered around the house and yard. If I took the Glow Ball away and instead threw a tennis ball, Cowboy would run to retrieve the ball, then return without it and begin looking all over the house for the Glow Ball. The Glow Ball is tough enough to survive Cowboy's big, sharp chompers, yet it's soft enough to do no harm when it hits him in the face. The glow feature is a huge bonus, too. Cowboy gets distracted by chipmunks and squirrels when we're playing, so he sometimes can't remember where he dropped the ball to pursue the interloper. If the ball was exposed to enough light before it was misplaced, it will be easy to find once the sun goes down. (Otherwise, Cowboy always finds it eventually.) When I'm feeling nocturnal, I'll put the ball under a light for a few minutes to activate the electrons, then treat my boy to a middle-of-the-night game of fetch.
I was so happy to see these on sale, I bought a lifetime supply for my pantry. The size guide seems accurate. ChuckIt recommended the large size for a dog of Cowboy's weight and that size is perfect. Highly recommended product.
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Reviewed in the United States on October 14, 2024
★★★★★ 5
Durable , dogs love them !
Size: 2 Count (Pack of 1), Style: Pack of 2, Size: 2 Count (Pack of 1), Style: Pack of 2
These chuck it balls are a fan favorite of my own dogs and my foster pups . So much so I have sent my foster home with them when they go to their new families! I seriously need an endless supply !
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Reviewed in the United States on April 30, 2026
★★★★★ 5
Whistles when thrown
Size: 2 Count (Pack of 1), Style: Pack of 2
One of my shepherd’s favorites! Truly does whistle when you throw it! Good quality!
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Whistle away my friend!
Size: 2 Count (Pack of 1), Style: Pack of 2
Love it! My dog loves it! The people at the dog park love it! The whistle is the best thing about these balls but the hard durable rubber and the denseness of the ball really make it fly. Let me quickly breakdown each of these:
First the sound is like a...well, whistle coming from a human but oscillating and slightly different. It helps the dogs track where it goes and helps them find it better when it is getting dark (while flying of course, not when it is stationary). It barely makes a sound when throwing it with the wind but really howls when throwing it into the wind. This is why you get this ball and most everyone at the dog park perks up and wonders what that is, asks about it, and thinks it is very cool; which it is!
Next is the durable rubber which doesn't have a single bite mark or split yet in 4 months of constant daily play for about 1 1/2 hours every day! The dog chews it as he brings it back and other dogs steal the ball and run with it while "killing" it perpetually and it has held up great. I don't claim this is going to be Kong ball durable but it is very well chosen rubber.
Last is the denseness of the ball which really makes to go far in a chuckit launcher. One doesn't really notice how far this goes until going back to a tennis ball and realizing how short a tennis ball goes compared to this. The whistler ball, and presumably other chuckit rubber balls (glow, irregular bounce), go 30-40% farther when I throw then using the longest chuckit thrower. This is excellent and wears my retriever out even faster; not to mention the rippling muscles he has from sprinting that whole way!
Overall I got both ball several months ago and we started with the blue ball and haven't had to touch the orange one yet. This is his goto ball at the park and it is the most popular ball, bar none, for all the dogs to steal. The bright blue aspect, the rubber aspect, the sound, and exclusivity of it make it coveted by dogs and owners. I might be gushing and glowing over this ball a bit much but I tend to do that when something unexpected ends up being part of your everyday life and blows you away at how wonderful it is.
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Reviewed in the United States on December 20, 2012