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Description
EdU-488 Cell Proliferation Assay Kit (6-well plate)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,
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 used for EdU experiments of about 200T in 6-well plates, and EdU experiments of about 2000T in 96-well plates or paraffin sections. The amount of use should be calculated according to the sample conditions during the experiment. I. Sample processing 1. Animal experiments (1) EdU injection in animals For mice, EdU can be prepared with PBS at a dose of 10 to 200mg/kg and injected intraperitoneally, locally into specific tissues or organs, 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 methods: according to customer experiments, such as intraperitoneal injection, subcutaneous injection, intramuscular injection, tail vein injection, etc., among which intraperitoneal injection is the most common. After 6 hours or according to the specific experiments to determine the appropriate time, quickly kill mice, take out the organization, in accordance with the general steps to make frozen section and paraffin section. EdU tag when also can consult relevant literature to adjust. 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 Before slicing processing: best cleaning tissues and organs, to remove residues in the tissues of the blood, EdU, reduce the background. It is advisable to slice thickness: 3-10 microns, biopsy had thick could affect the background and dyeing efficiency. Slice post-processing: Paraffin dewaxing: dimethyl benzene dewaxing 2 times, 15 minutes per times, gradient ethanol (100%, 95%, 85%, 75%), 5 minutes per times, deionized water elution 1 times. Processing of frozen sections: After 30 min at room temperature, the sections were fixed for 10 min and washed three times with PBS for 5 min each. 2, cell experiment (1) EdU labeling of cells a.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 100 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.Cells were washed 3 times with PBS for 5 minutes each time
(2) cells fixed fully 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: 488 chromogen = 855:40:100:5 (the reaction solution was prepared on the spot). add 50-500&mu per sample dropfold; L EdU dyeing liquid evenly cover samples (reaction), reaction liquid at room temperature away from light incubation 15 to 60 minutes, abandon the dyeing liquid reaction, PBS wash three times, each time for 5 minutes. < br / > 3, DAPI staining & have spent < br / > DAPI (100 x), with PBS according to the scale of 1:100 dilution into ready-to-use DAPI staining fluid, each sample in 50-500 & mu; L DAPI staining solution, staining in the dark for 15 min, and washing with PBS three times for 5 min each. < br / > 4, image photos & have spent < br / > dyeing advice immediately after the completion of observation, using fluorescence microscopy, confocal microscope or film scanner to capture images, after dyed dark glass slides 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." Compared with the traditional immunofluorescence staining (BrdU) detection method, EdU is only 1/500 of the size of BrdU antibody, and it is easier to diffuse in cells. It does not require strict sample denaturation (acid hydrolysis, pyolysis, and enzymatic hydrolysis), which effectively avoids sample damage and helps to observe the real situation of cell proliferation at the overall level of tissues and organs. Has higher sensitivity and faster detection speed.
According to PB, CU: AC: 488 chromogen = 855: spoken, Catalan than column configuration EdU reaction liquid (liquid of reaction is used). 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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| Storage Temp. | -20 ° C, valid for 12 months. |
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4.8 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Essential History for Retaining Reason and Rationality for Guiding the Nation
Awesome book on the History of people, themes and big money, corporate used in an attempt to make one religion the National Religion while neutering the First Amendment. It is true to it's title in detailing how Corporations and Religion came together to fight the New Deal of FDR. These two self serving entities fought for control of the people but did almost nothing to avert wide spread hunger and homelessness in the depression. The fortunes of corporations and insistence of religions on controlling the minds of people to produce jobs in their own occupations outweighed the immediate needs of the jobless and homeless. The opulent lifestyles of the big religions and their organizers came first over the health and welfare of the country, just like it had in Medieval Europe, where the Aristocrats and the Christian Churches ran the Continent. These two institutions were responsible for keeping Democracy out of Europe and ultimately creating the Soviet Revolutions in Russia that attempted to spread to the rest of the World. What is clear, from many books on this subject, that "Under God" means being subservient to God, the word of which comes from God's messengers. Subservience Under God means that when the opinions of the People differ from those of God as told through the Messenger, then God wins and the People Shut Up. Those that don't face stiff penalties. Fabulous History that everyone should know if they want to perpetuate a Democracy. The Roman Empire was "One Nation Under God", because the Christian Religion was the only legal Religion and it had only One God - no others were allowed and this was enforced with the sword. Rome will return to America if we are not careful.
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Reviewed in the United States on October 22, 2015
★★★★★ 3
Stranded on third base
Format: Hardcover
Prof. Kruse wanders away from the promise of the subtitle of his book--"How corporate America Invented Christian America." He almost gets there when he describes the ideological opposition of the wealthy to F.D.R's "New Deal." and their caricature of the "social gospel" as Communist propaganda. His description of Truman's encounter with Billy Graham is wonderful. For a book published in 2015, Kruse makes only slight mention of the fact that the politics of money and religion in contemporary times becomes more pernicious every day. The main body of the book drifts into redundant detail about "who prayed with who" and the spiritual influence of prominent evangelists on presidents from Eisenhower to Bush with only faint reference to their ties with the Capitalist agenda. He portrays wealthy Christians as true religious zealots when, in fact, they and many of the politicians they supported were likely hypocrites using religion as a wedge issue to increase their power and pursue their agenda of unregulated Capitalism. It is remarkable that Kruse entirely fails to mention the Koch brothers--billionaires who bankroll countless ultra right-wing politicians with evangelical agendas, while they, themselves, are apparently Ayn-Rand-style libertarian atheists interested only in a social-Darwinian, Capitalist heaven. Prof. Kruse leaves us stranded on third base, when he could have hit a home run!
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Reviewed in the United States on May 7, 2015
★★★★★ 5
Horrific Voter Suppression
Format: Paperback
This is a very well documented book about voter suppression in America. It was interesting to see that before the Jim Crow era, black people voted, were elected to office, got educations and did fairly well. With Jim Crow, horrific voter suppression began (lynchings, etc.). In the early days it was the Southern Democrats and the KKK. As time went on the methods simply became more devious. In the 1930s FDR began attempting to attract black voters to the party. By the end of WWII we had "Dixiecrats". By the time the time the Civil Rights Act was passed, the transition was complete and those Southern Democrats were now members of the GOP. (Much of that bit of history is not in the book, but you'll want to look it up.) The GOP is now the master of voter suppression, and you can see it pretty much everywhere but especially in Georgia, Texas, Minnesota and many southern states. The book is chock full or examples, case histories and stories of resistance. It made me want to sell my house and give the money to the ACLU, NAACP, and SPLC to defend our most precious right (not privilege) to vote.
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Reviewed in the United States on October 4, 2020
★★★★★ 5
This is a MUST read!!
Format: Kindle
This book is a must read! I was just not aware of the extent of the planned, organized, deliberate and insidious actions that the Republican Party has taken to suppress votes from the disenfranchised and the impact that has on our Democracy. I am stricken by the horror of what has been happening now for YEARS and is still going on today!! Carol Anderson has compiled hard data to demonstrate the pervasiveness of the many voter suppression steps that have been taken across many states, primarily in the deep south, but not exclusive to the south. It’s very difficult to take reams of data and put it into a coherent, logical sequence that can be easily understood but she is masterful at making her book eminently readable. This is a very data driven book but I had no trouble getting through it. The conclusion did have some hopeful information relaying steps that some states are taking to increase voter participation which did give me some hope. I am so glad that I read this and I think we all should be informed on this issue as it is central to our state as a country.
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Reviewed in the United States on April 30, 2021
★★★★★ 5
Great book but disappointing kindle experience
Format: Kindle
A must-read for anyone who wants to understand the troubled and racist history of voting rights in the US.
My only complaint is that the Kindle version of this book messes up the footnotes. The great thing about reading on the Kindle is that you don't have to flip back and forth to check the footnote cites; you only have to click on them. Sadly, the footnotes aren't correct in the Kindle version of the book. They somehow were shifted by one, as I was able to confirm in my hardcover copy, completely ruining the experience if you wish to read more about a specific incident. This is inexcusable in my mind, but I won't hurt the rating for this book by taking off a star.
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Reviewed in the United States on January 5, 2021