Has anyone tried to obtain recombinant IL-2 in HEK293 cells?

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How can I transfect HEK-293 cells in suspension without Freestyle medium?

Hi We were transfecting HEK-293 cells adapted to grow in suspension to produce recombinant proteins with excellent results. Cells were maintained in Freestyle, transfected with PEI/DNA also in...

10 November 2019 8,105 3 View

Can I use the Qiaprep 96 Turbo kit with a microplate centrifuge instead of vacuum?

I got from someone else Qiaprep 96 Turbo kits for high throuhghput plasmid DNA purification. But I dont have QIAVAC96 or a similar device to pass the solutions using vauum. Can I do it using a...

07 August 2018 1,830 0 View

How can I compare ELISA and BIAcore results?

Higher affinities result in higher ELISA signals, of course. But now Im working with several mutated ligands and their receptor. The wt ligand has an affinity to the receptor around 20nM. Several...

08 September 2015 7,119 8 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

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

Why activated CAR-Jurkat cell could not kill targets?

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

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

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

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

How to use Density Functional Theory to calculate carrier mobilities of solid system?

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

Does long-term culture of murine Monocytes result in macrophage differentiation?

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

How can I get my Granzyme B flow cytometry stain to be consistent?

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