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Description
Mitochondrial Membrane Potential Detection Kit (jc-10)Product Specification Usage 1. Preparation of JC 10 dyeing working solution: The amount of JC 10 staining working solution required for each well of the six well plate is 1 mL, and the amount of JC 10 staining working solution for other culture vessels is similar; For the cell suspension, 0. 5 mL of JC 10 staining working solution is required for every 500,000 to 1,000,000 cells. Take an appropriate amount of JC 10 (200 ) and dilute JC 10 in the
Product Specification
| Usage | 1. Preparation of JC-10 dyeing working solution: The amount of JC-10 staining working solution required for each well of the six-well plate is 1 mL, and the amount of JC-10 staining working solution for other culture vessels is similar; For the cell suspension, 0.5 mL of JC-10 staining working solution is required for every 500,000 to 1,000,000 cells. Take an appropriate amount of JC-10 (200 ×) and dilute JC-10 in the proportion of 8mL of ultrapure water per 50μL of JC-10 (200 ×). Thoroughly dissolve and mix JC-10 with vigorous shaking. Then add 2mL of JC-10 staining buffer (5 ×), and mix well to obtain JC-10 staining working solution. 2. Setting of positive control: CCCP (10 mM) provided in the kit is recommended to be added to the cell culture medium at a ratio of 1: 1000, diluted to 10 μM, and treated the cells for 20 minutes. Subsequently, JC-10 was loaded according to the following method, and mitochondrial membrane potential was detected. For most cells, the membrane potential of mitochondria will usually be completely lost after 20 minutes of 10 μM CCCP treatment, and it should show green fluorescence after JC-10 staining; However, normal cells should show red fluorescence after staining with JC-10. For specific cells, the concentration and duration of CCCP may be different, and it is necessary to refer to relevant literature to decide. 3. For suspension cells: (1) Take 100,000 to 600,000 cells and resuspend them in 0.5 mL cell culture medium. The cell culture medium can contain serum and phenol red. (2) Add 0.5 mL of JC-10 dyeing working solution, reverse it several times and mix well. Incubate in a cell incubator at 37 °C for 20 minutes. (3) During the incubation period, an appropriate amount of JC-10 staining buffer (1 ×) was prepared according to the ratio of adding 4 mL of distilled water per 1 mL of JC-10 staining buffer (5 ×), and placed in an ice bath. (4) After the incubation at 37 °C, 600g was centrifuged at 4 °C for 3-4 minutes to precipitate the cells. Discard the supernatant and be careful not to aspirate the cells as much as possible. (5) Wash twice with JC-10 staining buffer (1 ×): add 1 mL of JC-10 staining buffer (1 ×) to resuspend the cells, centrifuge 600 g at 4 °C for 3-4 minutes, pellet the cells, and discard the supernatant. Add 1 mL of JC-10 staining buffer (1 ×) to resuspend the cells, centrifuge 600g at 4 °C for 3-4 minutes, pellet the cells, and discard the supernatant. (6) After resuspending with an appropriate amount of JC-10 staining buffer (1 ×), observe with a fluorescence microscope or laser confocal microscope, or detect with a fluorescence spectrophotometer or analyze with a flow cytometer. 4. For adherent cells: Note: For adherent cells, if you want to use fluorescence spectrophotometer or flow cytometer detection, you can collect the cells first, and refer to the detection method of suspended cells after resuspension. (1) For one well of a six-well plate, aspirate the culture medium, wash the cells once with PBS or other appropriate solution if necessary according to the specific experiment, and add 1 mL of cell culture medium. The cell culture medium may contain serum and phenol red. (2) Add 1mL of JC-10 dyeing working solution and mix well. Incubate in a cell incubator at 37 °C for 20 minutes. (3) During the incubation period, an appropriate amount of JC-10 staining buffer (1 ×) was prepared according to the ratio of adding 4 mL of distilled water per 1 mL of JC-10 staining buffer (5 ×), and placed in an ice bath. (4) After the incubation at 37 °C, the supernatant was aspirated and washed twice with JC-10 staining buffer (1 ×). (5) 2 mL of cell culture medium is added, and the culture medium may contain serum and phenol red. (6) Observe under fluorescence microscope or laser confocal microscope. 5. For purified mitochondria: (1) The prepared JC-10 staining working solution was diluted 5 times with JC-10 staining buffer (1 ×). (2) 0.9 mL of 5-fold diluted JC-10 staining working solution was added with 0.1 mL of purified mitochondria with a total protein amount of 10-100 μg. (3) Detection with a fluorescence spectrophotometer or a fluorescence microplate reader: After mixing, directly use a fluorescence spectrophotometer to time scan (time scan). The excitation wavelength is 485nm and the emission wavelength is 590nm. If a fluorescent microplate reader is used and the excitation wavelength cannot be set to 485 nm, the excitation wavelength can be set in the range of 475 to 520 nm. In addition, fluorescence detection can also be performed with reference to the wavelength setting in step 6 below. (4) Observation with fluorescence microscope or laser confocal microscope: The method is the same as step 6 below. 6. Fluorescence observation and result analysis: When detecting JC-10 monomer, the excitation light can be set to 490nm and the emission light can be set to 530nm; When detecting the JC-10 polymer, the excitation light can be set to 525 nm and the emission light can be set to 590 nm. Note that it is not necessary to set the excitation light and the emission light at the maximum excitation wavelength and the maximum emission wavelength when measuring fluorescence here. For observation using a fluorescence microscope, when detecting JC-10 monomer, you can refer to the settings when observing other green fluorescence, such as the settings when observing GFP or FITC; When detecting JC-10 polymer, you can refer to the settings when observing other red fluorescence, such as propidium iodide or Cy3. The appearance of green fluorescence indicates that the mitochondrial membrane potential decreases, and the cell is probably in the early stage of apoptosis. The appearance of red fluorescence indicates that the mitochondrial membrane potential is relatively normal and the state of cells is also relatively normal. Precautions: 1. JC-10 (200 ×) will solidify and stick to the bottom, wall or cover of the centrifuge tube at lower temperatures such as 4 °C and ice bath. It can be incubated in a water bath at 20 ~ 25 °C for a while until it is completely melted. After use. 2. JC-10 (200 ×) must be fully dissolved and mixed with the ultrapure water provided by the kit before JC-10 staining buffer (5 ×) can be added. Do not prepare JC-10 staining buffer (1 ×) first and then add JC-10 (200 ×), as it will be difficult to fully dissolve JC-10, which will seriously affect subsequent detection. 3. When washing with JC-10 staining buffer (1 ×) after loading JC-10, keep the JC-10 staining buffer (1 ×) at about 4 ℃, and the washing effect at this time is better. 4. After the JC-10 probe is loaded and washed, try to complete the follow-up detection within 30 minutes. Store in ice bath before testing. 5. Please do not prepare all JC-10 staining buffer (5 ×) into JC-10 staining buffer (1 ×). JC-10 staining buffer (5 ×) should be directly used during the use of this kit. 6. If precipitate is found in JC-10 staining buffer (5 ×), it must be completely dissolved before use. To promote dissolution, it can be heated at 37 °C. 7. CCCP is a mitochondrial electron transport chain inhibitor, which is toxic. Please pay attention to careful protection. 8. For your safety and health, please wear a lab coat and disposable gloves. |
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| Beilstein | 0 | ||||||||||
| Description |
Mitochondrial membrane potential detection kit (JC-10) is a kit that uses JC-10 as a fluorescent probe to rapidly and sensitively detect the changes of mitochondrial membrane potential of cells, tissues or purified mitochondria. It can be used for early apoptosis detection.
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| PubChem CID | 0 | ||||||||||
| Storage Temp. | Dry and protected from light at-20 ℃, shelf life is 12 months. Ultrapure water and JC-10 staining buffer (5 ×) can also be stored at 4 °C. |
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4.8 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 3
Not indestructible but pretty darn tuff for sure!
Size: Large, Color: Blue
I would have maybe given a better rating if they hadn’t lied about it being indestructible and recommended it for my dogs size range! It didn’t last the first day. However my dog may be a bit above average when it comes to his relationship with the balls! It’s an issue and he should probably be in a 12 step program of some kind. But we all have our vices right and his is balls. All of them he needs em and he never stops joining for them and the stronger they are the more he’s driven to defeat it! So now that you have full disclosure from me. The ball is still in his box and it’s still round and mostly intact and that’s after several months! The part that didn’t last the first day was the squeaker! Poor little dude! If he would have just stayed quite it. But he didn’t and Bo dog knew he was in there! And made short work of him. Other than that. The ball pretty dang tough! But it’s not indestructible is it a big deal. Well in this world of way to many choices and then having to find the few that will tell you honestly what your getting and are not over stating their product! Just to get your money. And couldn’t care less about the quality! It sucks that we have to navigate all the crap to find the few decent things available. So it most certainly does matter! And I say that for their benefit the most hopefully! Because people are craving and constantly looking for a company that just makes the effort to put out the best they can. And tell you the honest description of what they have. That stands behind their work and puts more effort into quality then they do into profit. With the faith that the quality and integrity in their company will end up bring them the profits they can be proud of and know that they earned! Honestly! Is this company one of those! I don’t know? It’s not clear and in their defense I never reported the sneaky man’s demise on day one! I expected that. But indestructible is a descriptive word that has a very specific meaning. And when it’s used in a manner that it was! Then you’ve set yourself up to fail as there are very few things that meet that description! And I know I’m picking hairs here. But it’s words that get used like that wich makes one appear to have not met the standard they wanted to! And maybe you don’t realize its impact on your overall until it’s far down the road! Can it be said better. Sure . Always room for improvement. But I would certainly remove the claim that’s just unreasonable to expect. If I wanted to use the benifit the does have to offer perhaps I would have said this ball is the closest thing to indestructible we’ve made so far. That’s statement tell me that they are not claiming it is but that it’s pretty tough! And it with out exact words tells you these guys must take honesty seriously or they wouldn’t have chosen to claim the accurate step below. And that lets me know that they hold themselves to a standard that they obviously respect their customers and they don’t take them for granted that’s integrity right there and there’s just not enough of it o. This world today! Seems to me that there are a few that are really close to it though. Maybe so close it’s worth the time to just go ahead and say it. If my gut feeling is right then it’s absolutely worth the time! Do I recommend the item absolutely it’s as tough as nails! And if you hate the squeaky part. Then don’t worry it won’t last long if your dogs like mine. Or you can take the little dude out when the dogs sleeping! And I’m going to add a star because when I look at it in the wider view I’ve been a bit harder then is justified I believe. Their. Our lives today are not as good as they once were and should be! Because of our bad habit of not holding ourselves to the standards we are capable of and rather settling for what’s out there at the time. Because of many reasons right. What’s the point. Or it’s not going to change a thing. Or that as good as it needs to be. Don’t want to seem so picky and all the justification we inject in the place of our potential. That’s had a vey negative effect across the board! And we’re losing so much more then quality dog toys as a result. When we let or standards drop due to whatever reasons as a civilization then are standards drop. And that’s all you should need to know. To understand where that leads! And we only do ourselves the disservice and loss that comes from those choices. So look around and don’t hesitate to. All out that when you see it. Just try and come from a place that’s not to tear things down but to remind them of what they are really capable of! And how much we all need to hold ourselves to the highest standard we are capable of! And stop justifying excepting less! And then we see the results from that rather than the other.
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Reviewed in the United States on August 18, 2025
★★★★★ 5
MUST BUY! For large Dogs Safe & Fun! 100lb Dutch shepherd loves this
Size: Large, Color: Blue, Size: Large, Color: Blue
I have 100 pound Dutch Shepherd who likes to chew and he goes through toys pretty quickly. This one he has been chewing on for hours and has not made a dent, not one!! he loves it. He bounces it. It’s very bouncy. I love that about it the engagement with it is great! noise level. It makes some noise. I’ve got wood floors so you can hear it bouncing and the squeak is pretty loud, which is fine with me. It doesn’t bother me. My pet really really enjoys this a lot. I think this is an amazing value for the money because I’ve spent more money on other toys and they fell apart within a day. I think it’s a safe toy because he’s not got a little hard plastic pieces that are coming off of it. It does not break apart very easily and the size is good. it fits in the palm of my hand so it’s pretty good size ball for him. It’s big enough. He cannot swallow it, but he can fit it in his mouth. I think the thing that I like the most about it and he enjoys is that when he drops it, it bounces and he can chase it. Pet enjoyment 10 out of 10 engagement 10 out of 10. I feel it’s very safe dog friendly absolutely must buy hands-down. You will not be disappointed. It’s made of a very strong. I would say rubber and it’s super easy to clean. Just toss it in the sink. Wash it off with soap and water rinse it. It’s good to go. I’m running to buy another one right now as we speak so just in case he loses it, he’ll have a backup.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Great, solid toy safe for my dog
Size: Large, Color: Blue, Size: Large, Color: Blue
This ball is great! My pit bull love toys, especially ones with squeakers, but she’s a power chewer and trashes them almost immediately. Since she’s got a history of swallowing foreign bodies, this means she’s rarely been able to have toys, and when she does get them, it’s short lived. She LOVES this ball and it’s already lasted multiple weeks with no signs of any damage, despite the fact she carries it everywhere and chews on it daily. We don’t have to worry about her safety with this toy, because it’s large and tough enough she can’t swallow anything. It hasn’t lost its squeak either despite all her tough love. She loves to play with it and we’re getting more of them asap so she has a backup if this one gets lost or eventually does break.
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Reviewed in the United States on May 2, 2026
★★★★★ 5
Truly indestructible
Size: Large, Color: Blue
They are not kidding, this ball is truly indestructible. I've gotten several for my dogs so they have plenty to share. Two of my dogs are beyond super chewers (bully breeds). One of the balls is missing the section that's glued into most of them, but the structure has still held up. And my dogs LOVE them.
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Reviewed in the United States on May 31, 2026
★★★★★ 4
Tough STINKY ball
Size: Large, Color: Red
This ball is the only one I've found that my dog can not destroy in seconds. He can chew on it for a few weeks before he works a hole in it. That's the good part. The bad side is the horrible smell! It doesn't seem to bother him, but it is horrible to my nose. It is so awful that I tried soaking it and dish soap and vinegar water overnight to see if I could make it go away. It was a little bit better but still stunk. Anybody got other suggestions on what could make the smell go away?
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Reviewed in the United States on April 12, 2026
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