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Description
Mouse BCL2A1 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 uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with a Bcl2-related protein A1 (BCL2A1) 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 Bcl2-related protein A1 (BCL2A1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Bcl2 Related Protein A1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Bcl2-related protein A1 (BCL2A1) is a protein encoded by the BCL2A1 gene. This gene encodes a member of the Bcl2 protein family. Proteins in this family form heterodimers or homodimers and act as anti-apoptotic and pro-apoptotic regulators involved in various cellular activities, such as embryonic development, homeostasis, and tumorigenesis. The protein encoded by this gene can reduce the release of pro-apoptotic cytochrome c from mitochondria and block caspase activation. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Better than I expected!
Style: Variety Pack - Large (3-pack)
I have two pitbulls who can destroy any toy! My youngest one likes to chew on my fluffy slippers. She doesn't hurt them, just chews on the fur. I give her a lot of heck for this but she has an affinity for them. I decided to buy her a stuffed dog toy and she loved it! She proudly carried it with her everywhere and would throw it up in the air and catch it. She was as happy as she could be. The problem was that after a few days, my 5-year-old pittie wanted to get in on the action and they turned it into a tug of war. Five minutes later it was in pieces on the floor. I decided to see if there was such a thing as a cloth toy for aggressive chewers. I found these! They weren't that expensive, so I thought, "what the heck", I'll give them a try! They arrived the next day! I took two out of the pack and within two or three days they had killed the squeakers in both of them, and I was thankful! The squeaking was loud and they could not get enough of it. There were three squeakers in each toy! After the squeakers were gone, they seemed to lose interest. So I started doing tug of war with them. Pretty soon they were doing it with each other. I didn't expect them to last a day! However, it has been almost a month and they are still pretty much intact. I mean these dogs are for real and they pull as hard as they can and they still have not ripped them apart. And it's great exercise! There are a few small holes in them but they are not getting bigger. I guess that is how they got the squeakers out! Even the eyes on these toys are still there. I put the third one away for when these two are demolished, but I think I will order another pack of three. That will give me four all together! That should keep them busy for quite a while!
I just want everyone to know that I am pretty limited with my reviews. I don't bother unless I really have something to say, so you can totally believe what I am saying! I don't think you will be upset with this purchase, unless you don't like saliva soaked toys and/or your dog is prone to swallowing it's toys. It is very hard for me to imagine a dog swallowing one of these. From the top of their heads to the tip of their tails they are over a foot long!
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Reviewed in the United States on May 7, 2020
★★★★★ 4
Good quality. Decent price.
Style: Fox, Raccoon, Squirrel
The small size is perfect for my 13# dog, but I was somehow under the impression that one squeaker was in the head while the other was in the tail (and that might be the case for rhe larger size), but one us in the main body, with the other in the tail. The head is flat, and there's no way to push the sqeaker in the body up into the head.
Ordinarily, this wouldn't matter -- and my dog LOVES the one I gave him so far; he's squeaking up a storm! -- but I bought these toys specifically so I could fill the (presumably empty, but in reality not) body between head and tail with chopped up pool noodles so my paraplegic dog could take it into the water treadmill treatment he's starting in a couple weeks. I wanted something that would float and be largely waterproof (no fluff inside in case he drops it in the water), that he could hold in his mouth instead of wearing a muzzle.
I've rated these toys 4 stars because he does love them, they seem well made, and they're pretty affordable. I would buy them again, though not for the same purpose. (He hates rubber toys, so this was my last hope. Lol?)
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Reviewed in the United States on February 21, 2021
★★★★★ 5
Longest lasting dog toys we’ve purchased.
Style: Fox, Raccoon, Squirrel
In 2020, we purchased these for our 6 month old puppy who was repeatedly destroying all of her toys. Christmas 2025, I gifted our dog a stuffed toy purchased from a local dog store. It was destroyed within an hour. Later that day, I found one of these little squirrels that we purchased FIVE years ago - still in action, a bit worse for the wear, but still very much loved by our dog. I decided to purchase another pack of these as they are clearly winners.
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Reviewed in the United States on January 19, 2026
★★★★★ 5
Nice toy
Style: Variety Pack - Large (3-pack)
Good idea, no stuffing.
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Reviewed in the United States on May 9, 2026
★★★★★ 3
The Ad got me, the reality didn't
Style: Variety Pack - Large (3-pack)
Directly from the ad- "Exclusive Noisemakers - Each large toy includes 3 high-quality round squeakers to deliver more sound to keep your best friend entertained.". Now my neighbor bought this squirrel with three high-quality squeakers nearly a year ago. It's ragged and tattered I just recently sewed it up for him where he chewed the one leg odd, but those three noise makers are still squeaking. Whe I got my pup, I immediately went to buy one and they were no longer selling them, so I've been on a search for one or an equivalent. These are not.
There's no stuffing to get all over the place when your pup breaks into it, a plus. However the squeakers weren't particularly loud and were silenced in a few hours. Two were dead and the remaining one was very well on its way. The toy itself was pretty much intact but that had more to do with the lack of attention from the dog than how rugged the toy is. He'll pick it up once in a while but it certainly didn't meet my expectations.
The good thing is that there are three of them so I can give him some brief excitement in the future. Wouldn't buy again.
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Reviewed in the United States on February 24, 2023
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