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Description
EdU-555 cell proliferation assay kitProduct Specification Usage Before the trial guidelines 1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg kg (50 mu g g), dilute concentration is 0. 5 1 mg mL. Are injected into mice weight 20 g, mg, 1 to 1 mg mL, need injection of 1 mL. 2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different,
Product Specification
| Usage | Before the trial guidelines 1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg/kg (50 mu g/g), dilute concentration is 0.5 1 mg/mL. Are injected into mice weight 20 g, mg, 1 to 1 mg/mL, need injection of 1 mL. 2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different, can be adjusted appropriately. 3, this kit can be carried out about 200 t of 6 orifice EdU experiments, about 2000 t96 orifice or paraffin section EdU experiment, experiment should be calculated according to the sample situation in the process of usage. I. Sample processing 1, the animal experiment (1) EdU injection in animals In mice, can according to the amount of 10-200 mg/kg, the EdU with PBS mixed with a certain concentration, intraperitoneal injection, specific tissues or organs local injections or added to drinking water. The specific dosage is related to the type, weight and mode of use of animals, and can be referred to the relevant literature. Therefore, it is recommended to conduct a certain exploration of the use concentration of EdU for the first use, or directly use the concentration of 50mg/kg for testing. Injection method: the experiment according to the customer, such as abdominal cavity injection, subcutaneous injection, intramuscular, tail intravenous injection, etc., among them with intraperitoneal injection. After 6 h or after the appropriate time was determined according to the specific experiment, the mice were quickly sacrificed, the desired tissue was removed, and frozen or paraffin sections were made according to routine procedures. EdU labeling can also be adjusted by referring to the relevant literature. Small intestinal epithelial cells proliferate rapidly, and positive information can be detected 6 hours after EdU injection in adult mice, which can be used as a positive control for pre-experiments. (2) Slice processing Pre-section treatment: It is best for tissues and organs to be washed to remove residual EdU in blood and tissues and reduce background. Section thickness: 3-10μm is appropriate, too thick section may affect the section background and staining efficiency. Slice post-processing: Paraffin sections were deparaffinized twice with xylene for 15 min, once each with gradient ethanol (100%, 95%, 85%, 75%) for 5 min, and once with deionized water for elution. Frozen section processing: room temperature placed after 30 minutes, fixed 10 minutes, PBS cleaning three times, each time for 5 minutes. 2. Cell experiments (1) cells EdU markers a.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 10 microns EdU medium; b.each hole adding suitable amount of 10 microns EdU medium 2 hours incubation, abandon culture medium (optimal incubation time for 1/10 of the cell cycle) commonly, EdU incubation medium and EdU reaction liquid volume may refer to the table below; c.PBS cleaning cell 3 times, each time for 5 minutes.
a. Cell fixative (PBS containing 4% paraformaldehyde) was added to each well and incubated at room temperature for 15 minutes. The fixative was discarded and washed with PBS for 3 times, 5 minutes each time. b. Each well was decolorized with an osmotic agent (0.5% TritonX-100 in PBS) and incubated for 15 minutes. PBS was washed three times for 5 min each. Second, EdU response
The EdU reaction solution was prepared according to the ratio of PB: CU: AC: 555 chromogen = 855:40:100:5 (the reaction solution was prepared immediately). Each sample add 50-500 mu L EdU staining reaction liquid liquid evenly cover samples (reaction), 15 to 60 minutes at room temperature away from light incubation, abandon the dyeing liquid reaction, PBS wash 3 times, each time for 5 minutes. 3. DAPI staining DAPI(100x) was diluted with PBS at a ratio of 1:100 to make ready-to-use DAPI staining solution. 50-500μL of DAPI staining solution was added to each sample and stained in the dark for 15 minutes, followed by three washes with PBS for 5 minutes each. 4. Take pictures Fluorescence microscope, confocal microscope or whole-film scanner were used to collect images. After staining, the slides were stored in a dark place at 4℃. Above are stained images of 50mg/kg labeled mice for 6 hours |
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| Description | EdU(5-ethynyl-2' -deoxyuridine), also known as 5-ethynyl-2' -deoxyuridine (5-ethynyl-2' -deoxyuridine), is a thymidine analogue with an alkynyl hydroxyl group that is rare in natural compounds and can replace thymidine (thymidine, 5-ethynyl-2' -deoxyuridine) in cell proliferation. thymidine (thymidine) is inserted into the replicating DNA molecule, the ethynyl group on EdU can react covalently with the fluorescently labeled small molecule Azide probe (Azide Alexa Fluor 488, etc.) through the catalysis of copper monovalent ion to form a stable triazole ring. "It is called the Clickreaction, which allows for an efficient and rapid measurement of cell proliferation, especially the percentage of cells in S phase." With traditional immunofluorescence staining (BrdU) method compared with EdU only 1/500 the size of the BrdU antibody, are more likely to spread within the cell, don't need strict sample degeneration (acid solution, pyrolysis, enzyme solution) processing, effectively avoid the damage samples, help in the overall level of tissue and organ observation of cell proliferation, It has higher sensitivity and faster detection speed.Note: AC is a powder, need to add 10mL pure water before use, fully dissolved before use, AC is easy to oxidation failure, after the refrigerator is stored at -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg/mL, which should be converted according to the dosage.
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| Storage Temp. | -20 ° C, valid for 12 months. |
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4.3 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
Best dog toy!
Color: Air Head, Size: Large Dog, Color: Air Head, Size: Large Dog
I got this toy for my dog who loves squeakers not only is it a two in one it actually has three toys inside which I think is amazing!! 10/10 recommend
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Reviewed in the United States on June 11, 2026
★★★★★ 5
Love all the Barkbox toys
Color: Christmas Consuela, Size: Large Dog
This is our third "Consuela"! Our dogs LOVE these toys, and they are the most durable we can find for our heavy chewers. They really like the ball inside as well. Highly, highly recommend these toys. I won't be buying other toys anymore, just barkbox.
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Reviewed in the United States on March 27, 2026
★★★★★ 5
My dog is a toy destroyer
Color: Consuela the Cactus - Large, Size: Large Dog, Color: Consuela the Cactus - Large, Size: Large Dog
I’ve had a hard time trying to find toys that my dog, Zara loves because she’s actually very picky. Ive gotten plenty of Kong toys that are well known to be indestructible but she wants nothing to do with them. Then I got her ropes and that seemed to be good for a while until she was getting sick in the middle of the night from eating all of the string. Stuffed toys aren’t good because she’ll rip them within seconds and eat all of the stuffing. I felt like I’ll never score the perfect toy for her.
I found the three layered cactus toy and decided to give it a go. It’s clearly designed to be ripped and destroyed to get the ball in the middle which is actually really cute and fun to see all of the layers.
The cactus arms have stuffing and of course that’s the first thing she went for and ate. I had to watch her and make sure I took the stuffing away before she ate it. And before I knew it, she ate half of the first layer fabric lol once she got through the first layer, all she’s been doing is walking around the house with the toy in her mouth. But now she’s tired and passed out on the couch from all of the fun.
This toy won’t last long with a toy destroyer but it sure does give them something to do and it’s a cute toy. I do recommend!!
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Reviewed in the United States on July 26, 2024
★★★★★ 5
Last a while. Will be buying more toys soon.
Color: Consuela the Cactus - Large, Size: Large Dog
Best dog toy I have ever bought. I have three dogs, two at the time I bought this toy. It took them over a year to break through the first layer. The flower comes off first so be careful your dog doesn't swallow it. It still has the second layer so it still looks like a cactus. No stuffing to clean up when they tear into it, which makes it far less messy than other toys. One of my dogs is obsessed with fetch so I use it like a ball inside. The oval shape makes it hard to ensure when it bounces that it will bounce straight. It's bounced into walls and other stuff that I wasn't intending it to hit. Not much of a squeak so I can let them play with it for hours and I don't get annoyed with the noise. Great toy that lasts a while.
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Reviewed in the United States on September 25, 2025
★★★★★ 5
Nice Value
Color: Consuela the Cactus - Large, Size: Large Dog
Very cute! Its soft and feels like it is well made. It is a heavier toy and a tougher one. Not a plush toy. I will order again for our older dogs. Nice value for sure!
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Reviewed in the United States on June 11, 2026