Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 7 - Aug 12
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human CASP3 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, 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. 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. |
|||||||||||||||||||||||||||||||||
| 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 Caspase 3 (CASP3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Caspase 3 (CASP3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Caspase 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Caspase-3 is a caspase protein that interacts with caspase-8 and caspase-9. It is encoded by the CASP3 gene. CASP3 homologs have been identified in many mammalian species for which complete genomic data are available. CASP3 is a member of the cysteine-aspartate protease (caspase) family. The sequential activation of caspases plays a central role in the execution phase of apoptosis. It exists as an inactive proenzyme that undergoes proteolytic processing at a conserved aspartate residue to produce two subunits, a large and a small, which dimerize to form the active enzyme. This protein cleaves and activates caspases 6 and 7, which are themselves processed and activated by caspases 8, 9, and 10. It is the primary caspase involved in cleaving the amyloid β-4A precursor protein. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.1 ★★★★★
Based on 13 reviews
Sort
Product Reviews
★★★★★ 5
Recommend for my pit mix.
Size: Large, Color: Blue, Size: Large, Color: Blue
Wasn't sure how this leash was going to be for my dog. He is a pit/border collie mix and I usually have a harness on him.
No worries, he was great! Let's me put it on, responds to corrections and is much easier to handle. Why didn't I buy this earlier?
Highly recommend.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 15, 2026
★★★★★ 5
Flexible and secure for your dog
Size: 48" One Size Fits Most, Color: Black
Excellent quality and durability. The design is simple, flexible, and easy to use. When used properly, it works well without choking and feels secure and comfortable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 25, 2026
★★★★★ 3
Does help, but needs constant adjusting during walk
I watch so many Cesar television shows on how to correct dog behavior and see him use a Yellow version on the shows. I see there is not a yellow version of this lead available on the site so that was irritating because I wanted to get what he uses exactly from his show. Anyway, thanks to another reviewer, I knew to buy the "large" size of this for my 40lb dog because she made the mistake of buying the regular smaller size this site defaults to choose. The "large" is not very long, only 2 ft but maybe the rope is thicker.
The rope is a light stretchy material so it does not chafe your hand, but it still burns after a while of your dog pulling and you trying to keep it from pulling. I could not imagine trying to use this on a dog bigger than 40 pounds because it feels like it is at it's max strength when I am dealing with mine.
My dog likes to eat anything and everything on the ground as we walk so I needed this for the option of the muzzle control feature of the rope. As others have said it acts like the gentle leader halter.
Pros: The adjustable slide to "secure" the loop size over your dog's head, behind it's ears is easy to move up and down.
The black metal ring works well when you use the second circle to loop over your dog's muzzle. Every time she tries to open her mouth to eat something on the ground she can't.
CONS: The adjustable slide to "secure" the loop is too easy to move and therefore I have to constantly push it back down on my dog's head while walking. I am trying to find a clamp of some sort I can clip behind this slider to keep it in place once my dog starts moving.
As soon as it begins to slide up from movement, causing the neck hoop opening to widen and she is back to pulling hard.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 7, 2022
★★★★★ 5
Such a great tool!
Size: 48" One Size Fits Most, Color: Black
I have had this product for a week, and I have nothing but good things to say about it! I debated for a couple of days as to whether or not to purchase it because the price seemed a tiny bit much for what looked like such a simple item. Now I know it’s that very same simplicity that makes this such an effective tool! Best money I could have spent on my dog!
I’ve had my rescue Patterdale Terrier for a little under 3 months and I have quickly found out he is far from being perfect. It has taken a lot of time to begin working on correcting some of his problems, one at a time. Like many dogs, he also pulls when being walked. So what did I do as a regular person in need of quick dog tips? I searched up Cesar Millan videos on YouTube. 😂
I had purchased a Flexi leash two weeks after adopting my dog, and he broke it within another two weeks. While it was disappointing when I first found out, I now see it as a blessing in disguise. Paying close attention on YouTube, I noticed common features in all of the videos where the owners had pulling problems. Most owners in these videos had Flexi leashes, and the dog is always walking in front of the owner, not besides them. All Cesar uses to start making a change are leads just like this one. He never explicitly promotes his lead on his show, but after seeing the immediate increased control using such a simple tool, I decided to look online and found this. So I figured, why not?
My dog is 28lbs, so he isn’t big enough to overpower and drag me during a walk. After the Flexi broke, I resorted to using a regular hook-through-the-collar leash. The pulling problem was still there, and would only get worst if we came across a squirrel. As soon as we started using this lead, he’s become much more responsive to my corrections. As soon as he is too far in front of me, even without actually pulling, I give it a quick, gentle tug and a “Chh!” and he‘s learned that he needs to slow down. If he actually pulls and won’t immediately respond to my correction, we’ll sometimes stop walking altogether until he sits, calms down, and realizes that we’ll only walk at my pace. This lead has helped tremendously and there’s rarely ever any tension on the lead when walking now. It’s hard to believe it has only been a week since I’ve started using it, and I don’t believe I’ll go back to using the traditional leash unless absolutely necessary. I would definitely purchase this again!
And no, I have not been paid by anyone, nor was I sent this by any company to review it. Just a regular girl who is so happy to have found something that is actually seeming to work!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 4, 2018
★★★★★ 5
Worked from day 1
Size: 48" One Size Fits Most, Color: Black
This leash is the best thing I’ve ever bought for my dog. He’s 1 year and 4 months old, and I’ve bought at least five different harnesses since I got him, including very expensive ones like DogfriendlyCo. He pulls with every single one of them.
I bought this leash a while ago, but I only used it once because I was afraid he would get hyperactive and choke himself. He hated it at first and tried really hard to get out of it. Today, after four months, I decided to give it another try and omg! This is the first time I’ve been able to walk him without feeling anxious or embarrassed. He walked by my side the whole time, and the leash stayed loose. Iwould only pull a little when he wanted to go ahead of me. I could tell he enjoyed the walk and so did I.
I’ve tried it a couple more times just to make sure it wasn’t luck, but this thing really works. Highly recommend.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 23, 2026
recommand products
Sexy Sheer Mesh Bikini Set
35.00
Shanghai deluxe edition Map, Olivier Gratton-Gagne
124.50
My Favorite People Call Me Gabriela , Black, Women's Short Sleeve Round Neck T-shirt, gift t-shirt 00371
20.00
Leopard Print One Piece
35.00
Men's Graphic T-Shirt Abuja Limited Edition Short Sleeve Tee-Shirt Vintage Birthday Gift Novelty
20.00