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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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4.8 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
I use it for myself not my pet
I sewed a fluffy velcro pouch to put this in, and then I wrapped the pouch around a microwavable heat gel pack in order to get the sensation of both heat and heartbeat while hugging a large stuffed animal. It works great. It's not too loud if you position it right. It helps me to fall asleep and not feel lonely or touch starved. The battery lasts a long time and I don't have to recharge it very often. I only wish that the speed of the heartbeat was adjustable so that I could slow it down a little more so that my own heartbeat might try to match it.
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Reviewed in the United States on December 7, 2024
★★★★★ 1
Easily Exposed Lithium Ion Battery
I am returning this item for my money back. Apparently, they think they can charge $16 for a piece of plastic and not even put a screw on the back to hold the panel closed. My dog is smart and a little rough when anxious. I wanted a rechargeable heart (to avoid buying batteries) to help with this anxiety but cannot risk the Velcro coming undone and him biting the heart by mistake. The panel slides off very easily. There's a reason that lithium ion batteries aren't usually easily accessible. I'm not risking my dog discovering a battery and wire in the heart. Even with appropriate supervision, accidents happen. I don't even understand why I would need to access the battery when it's rechargeable. I don't recommend purchasing this product for that reason alone.
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Reviewed in the United States on July 17, 2022
★★★★★ 4
Works great!
It works great, don't have to buy batteries.
07/-8/25
Marilyn A.
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Reviewed in the United States on July 9, 2025
★★★★★ 5
bottle baby kittens preferred this heartbeat to having warm snuggle safe heat disk
Ok... when I got this I thought.. "are you kidding me?" they are going to like this.
Joke was on me. They love it. It is rhythmic and it is strong enough that it pulsates the area so all of them can feel it. They consistently would lay on top of the heart rather than in the area of their "crib" where the heat was.. and this was when they were still only a few days old! It works.
So this is what you want to know.
It beats continuously for about 4-5 days without needing a recharge.
Recharge is fast and only takes a couple hours
The button is slightly "inside" the unit so when it is placed upside down there is no way that it can accidentally be turned off. Which is great. If the button were on the upside in the pet space then a foot or body part of the little animal could turn it off inadvertantly. Design to put the button slightly inset is extremely important design feature - makes this excellent. When I turn it on I know it will be going as long as the battery still has charge. .
I don't think it is waterproof so I put it under the pee pad that I had under the blanket bedding and it was still easily heard AND felt. If not using a pee pad I would definitely put it in a small ziplock bag to protect from any leaked urine or liquids to keep it safe.
When the kittens were done eating they would gravitate towards the heart beat... (with their eyes open or when they were still closed after birth it was the same response). This little gadget was a total lifesaver as it really calmed the kittens in ways I would not be able to.
These pictures are of the trio that LOVED this.... Meet Zen (tabby M with white feet), Sugar (tuxedo F with white feet) and Rocket (ginger - M). All cisgender. they were about 3 days old when I got them to foster.. photos from arrival until about 3 weeks of age. Photo with q-tip is to give relative size on arrival. They will be up for adoption in Tustin CA in another 2-3 weeks time.
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Reviewed in the United States on September 17, 2023
★★★★★ 5
Fantastic!
The heart that came with Snuggle Puppy was a pain to replace the battery. This is the only rechargeable heart I could find searching online. I was a little concerned reading it’s a “frequently returned item.” But happy to say, we will not be returning this! It shipped charged and the heart is still beating non-stop days later! It’s sturdy and fits in the snuggle puppy perfectly. We have a tiny 17 yr old Chihuahua who never chewed anything up so it’s completely safe for her to enjoy. It helps with her anxiety. You can hear the beat in the video—it’s nice and loud and has a bit of a vibration. Our pup has been sleeping next to it every night. Very happy with this purchase. From reading reviews, I trust if we have an issue with it, their customer service will help. For now, it works! I really like that it has time options—can set it for 2 hrs of heart beating or non-stop.
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Reviewed in the United States on September 17, 2024