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Description
EdU-594 cell proliferation detection 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: 594 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 12 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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4.1 ★★★★★
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Product Reviews
★★★★★ 5
A Beta Worth Rooting For
Format: Kindle
In Spare, Violet Fox flips the omegaverse on its head, giving us a Beta heroine determined to make her mark. Joining the Beta Trials to support her sick father, she's thrown into a pack that doesn't want her, especially the possessive Alphas.
But here's the twist: their sweet Omega turns out to be her scent match. Cue the angst, forbidden tension, and a slow-burn romance that will make your heart ache in the best way.
Violet Fox delivers an emotional, refreshing take on the genre, proving Betas aren't "spares." They're stars.
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Reviewed in the United States on January 10, 2025
★★★★★ 5
Beta, Alpha, Omega oh my!
Format: Kindle
Omegas are precious and given to Alphas & their packs... but the Betas want in too. To this end, the Beta government is rolling out its trial of assigning a Beta to each Alpha-Omega pack. But forcing a Beta into a pack where they are not wanted will not end well... Of course, no one expected the Omega to fall for the assigned Beta. Great read and cliffhanger
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Reviewed in the United States on February 15, 2025
★★★★★ 3
A familiar story, just with…..less.
Format: Kindle
So, as other reviewers make clear, this is very similar to Pack Darling and The Beta. It’s much closer aligned with The Beta, in plot and maybe more like Pack Darling with characters.
That being said, I don’t hate this…..but it wasn’t great either. It’s both books mentioned but just….less. Less angst, less emotion, less feeling. The plot feels very half fleshed out, and the “bad guy” feels underwhelming. I didn’t really feel any real emotions from and of the male leads, except maybe Oliver. The others fell sorta flat for me. And Mika makes herself out to be this big bad ass straight outta training and then we never see it from here again with the one fitting room incident as the exception.
SPOILER:
The whole, “Oh, I’m actually probably an Omega, but I don’t wanna be but I do actually wanna be but no one can ever know my secret that I do nothing to hide “ thing fell so flat. She never commutes to believing she was secretly an omega, but also mentions her “secret” a lot. It just felt so manufactured.
I’m intrigued enough to read part 2 and see how the author closes everything out, but this is not one I’ll recommend or ever come back to.
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Reviewed in the United States on February 13, 2024
★★★★★ 5
A good read
Format: Kindle
A good read, just fluffy cuteness, no antagonism. I like all the characters. It could have used another round of editing however, glanfds being one error that cracked me up, and my personal pet peeve was that the author kept using the word fill instead of feel, which I promise you are not interchangeable haha, but it's definitely better than the majority of books I read on here mistake-wise.
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Reviewed in the United States on November 28, 2024
★★★★★ 4
amazing
Format: Kindle
Knot the Bride was a fantastic read! The characters were all amazing and well-developed. It was easy to like them all. Sophia, Luca, Nick, and Gavin were all perfect for each other. It was such a charming story that had me hooked the entire time. I did wish there were POVs from Luca, Nick, and Gavin but it was still an amazing book without it.
I am excited to read the next book in the Willowside Omegaverse series! This is definitely a must-read for fans of omegaverse romance!.
I received a free copy of this book and am voluntarily leaving a review.
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Reviewed in the United States on April 25, 2025