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Description
Mouse Liver Organoid Culture Medium KitProduct Specification Usage 1. Primary (1) the tissue after sampling must be placed in the pre cooled (2 8 c) sampling bottle of tissue preservation solution e, and quickly transferred to the clean laboratory for tissue processing and cell separation, taking photos and registering information (2) prepare several Petri dishes and add 4 pre cooled primary culture buffer B for standby (3) disinfect the sampling bottle, put the tissue into the Petri dish,
Product Specification
| Usage |
1. Primary (1) the tissue after sampling must be placed in the pre cooled (2-8° c) sampling bottle of tissue preservation solution e, and quickly transferred to the clean laboratory for tissue processing and cell separation, taking photos and registering information (2) prepare several Petri dishes and add 4 ℃ pre cooled primary culture buffer B for standby (3) disinfect the sampling bottle, put the tissue into the Petri dish, wash it three times with primary culture buffer B, remove impurities, and cut the tissue into a tissue block with a volume of about 1-3mm3by using ophthalmic scissors or scalpel (4) the tissue was digested with the digestive solution c of primary mouse normal liver tissue, and then it was shaken at 37 ℃ for 10-20min (the digestion condition was observed at any time during the digestion process) (5) take a small amount of liquid and observe it under the microscope. After observing more single cells or cell clusters below 70UM under the microscope, add three times the volume of primary culture buffer B to stop digestion (6) filter with a 100um pore size screen, collect the filtrate, remove the supernatant after centrifugation at 300g for 5min, add primary culture buffer B and resuspend for centrifugation (7) Matrigel calculation: after step 6, observe the collected tissue volume, add 25 times the tissue volume of Matrigel (abs9495) to resuspend the plate (8) take a 24 well cell culture plate as an example, dispense 25ul of tissue Matrigel mixture per well for plating (operate at 4 ℃) (9) put the laid culture plate into a 37 ℃ incubator for 10-15min to form a gel, and add mouse normal liver organoid medium a (restore room temperature) for culture 2. Organoid subculture (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) a: the number of organoids is insufficient or the volume is small: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4  &Nbsp&Nbsp&Nbsp b: When the number of organoids is large or the volume is large: centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid passage digest D for digestion for 2-3min, add organoid passage culture buffer g to stop digestion, centrifuge for 5min to discard the mixture, add an appropriate amount of organoid passage culture buffer g to resuspend and transfer into a 1.5ml centrifuge tube, centrifuge for 5min at 300g to discard the liquid for step 4 (4) after organoids were collected, add Matrigel to resuspend, lay 25ul Matrigel per well in a 24 well cell culture plate, place in the incubator for 10-15min, and add 500ul of mouse normal liver organoid culture medium a 3. Organoid cryopreservation (1) pipette the culture medium, add 1-2ml of 4 ℃ organoid subculture buffer g to each well and place it for 2min (2) pipette the Matrigel gently, collect it in a 15ml centrifuge tube, and let it stand at 4 ℃ for 10min. (each 6-8 well is a group) (3) centrifuge for 5min to discard the supernatant, add an appropriate amount of organoid subculture buffer g to resuspend, and centrifuge for 5min at 300g to discard the liquid (4) add an appropriate amount of organoid cryopreservation solution F, gently blow and resuspend. Take a 24 well cell culture plate as an example: the density is 2 wells and 1 tube is frozen, with a volume of 1.4ml per tube (5) mark the information, and after cooling down the program, transfer it to liquid nitrogen for long-term storage 4. Organoid resuscitation (1) take 10ml of organoid subculture buffer g and put it into a 15ml centrifuge tube (2) take out the frozen organoid cells from the liquid nitrogen tank and quickly put them into a 37 ℃ water bath pot for melting (3) during the water bath thawing process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely thawed in 1-2min (4) quickly transfer the dissolved organoid cells to a 15ml centrifuge tube, gently blow 6-8 times with a pipette, centrifuge at 300g for 5min, and then remove the supernatant and collect the organoid cell pellet. Add an appropriate amount of organoid subculture buffer g to resuspend and transfer into a 1.5ml centrifuge tube. Centrifuge for 5min at 300g (5) resuspend Matrigel, lay 25ul Matrigel per well in a 24 well cell culture plate, place in an incubator for 10-15min to form a gel, and add 500ul of mouse normal liver organoid culture medium A. |
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| Description |
Composition:
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| Storage Temp. | 4° C. 3months; -20° C. 1 year, see label for details |
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4.4 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 4
not bad
Format: Kindle
I loved the plot of this book. The characters just didn’t have a lot of depth. The connections and “love” just weren’t communicated very well in the writing. The author didn’t write the sweet psycho trope very well at all either. Lachlan was just a mess of a character.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 17, 2023
★★★★★ 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.
WAS THIS REVIEW HELPFUL?YesReportShare
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
WAS THIS REVIEW HELPFUL?YesReportShare
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
Good start to a series
Format: Kindle
I delayed reading the series for reasons I don’t remember. But my TBR list is huge so I thought I’d take a shot of this and I was pleasantly surprised. I didn’t think the blurb about it was anything special. But it was a very good book. It took some interesting twists and turns. I am so glad the second book is already out. Because I would not have waited patiently. Very slow burn but good storyline. 🔥🔥/5
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Reviewed in the United States on January 3, 2025
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