I am wondering if Ig-kappa leader sequence (METDTLLLWVLLLWVPGSTG) would allow my protein to be secreted into the media in hepatocyte cell lines: HepG2 or Huh7? Has anyone tried it? Did your protein got secreted?
Dear everyone, I work to improve the function of IRESes to make translation of mRNA and circRNA more efficient. The IRESes we are using are not very strong. I already looked at IRESite. If you...
15 February 2023 1,281 2 View
Dear colleagues, Could you please suggest the best way to thaw and freeze purified mRNA in water. I have batches of unencapsulated mRNA in nuclease-free water supernatant from nanoparticles and...
09 August 2022 3,269 3 View
Dear colleagues, I have applied 1 ug/mL of doxycyline to human iPSCs but most cells died within 24 hours after treatment. I was wondering if you have applied doxycyclin to iPSCs and know what...
26 January 2022 1,459 1 View
Dear all, I am planning to study histology and want to do that at home. I have only stained a blood smear with Giemsa but now want to try a wider range of animal tissues. What reagents will I...
24 February 2021 5,919 3 View
Dear all, I am trying to harvest the low-expression plasmids in the mentioned E.coli strain. I constantly gel very low concentrations (30-50 ng/uL while the concentration is usually around 100...
08 February 2021 4,468 4 View
Dear all, I am propagating some plasmids with a human gene and an ampicillin resistance gene. My logic is that if ampicillin concentration will be increased, the concentration of plasmid (pSF)...
14 January 2021 992 3 View
Dear all, I was doing site-directed mutagenesis with one mutagenesis primer (fwd) and one normal primer (rev) to amplify a 6.5-kb plasmid. Then KLD was performed with dnp1 destroying the original...
10 January 2021 1,197 6 View
Good afternoon, I have stored P2 and N3 buffers in ice for about one hour. The P2 buffer turned milky and I discarded it. The N3 buffer didn't seem to have any chemical changes, so I am wondering...
24 December 2020 6,020 0 View
Dear everyone, May you please recommend a protocol for electroporation of NK-92 cells using Neon machine? I have never done that before and woul dbe happy for recommendations if anyone had...
01 January 1970 1,856 0 View
Hi everyone, What statistical test would be most appropriate for this experiment? I want to compare two conditions against one another at different time points, e.g. mRNA1 vs mRNA2 at 24 hours,...
01 January 1970 3,102 5 View
I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...
07 August 2024 7,535 4 View
Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...
06 August 2024 641 2 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 View
Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...
04 August 2024 8,894 1 View
Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...
04 August 2024 7,282 2 View
I have used PE and PE-Dazzle 594 fluorochromes and have managed to get NK cells to properly show GranzymeB expression after 4 hr PMA/ionomycin stimulaton, but for some reason my CD8 cells in the...
01 August 2024 7,677 2 View
What types of mask designs for metal deposition, to be used in a PVD system, are best suited for perovskite-based solar cells?
31 July 2024 4,835 3 View
Hi there! I recently started working on Lx-2 cells. When I thaw the cells, they start attaching within few hours but later in about 2 days they started rounding and dying. I used DMEM+2% FBS...
31 July 2024 3,893 0 View
Hi all, may I ask you for advice about treating monolayer culture cell line with essential oil. Due to the hydrophobic character of the oil, I am concerning how can it equally affect to all the...
31 July 2024 5,301 2 View
As animals learn a task, they become more reliant on their long-term memories as compared to the real-time sensory information to guide behavioral performance (Ahilan et al. 2018). This process...
31 July 2024 9,859 0 View