What protocol is most to transfect lung endothelial cells (mLECs) with plasmid?

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A variable moderating multiple relation - how can I test hypotheses using amos?

How can I analyse the relationship between five independent variables and one dependent variable, with a moderating variable influencing each of these five relationships? How can I test hypotheses...

03 June 2024 6,221 1 View

Any experience in fibroblast centrifugation?

We are experiencing different problems pelleting fibroblast: a) From the resuspension of the comercial vials (after defrosting, we resuspended in 5ml complete DMEM + L-Glutamin + FBS +...

28 May 2024 3,529 1 View

Fibroblast trypsinization in Hanks medium with Ca and Mg?

We are setting up fibroblast cultures in a new location and we had to change the mediums suppliers. At the trypsinization step trypsin did not work and we realized our hanks medium has Ca and Mg....

28 May 2024 6,052 2 View

Are the calibration curves of NO radical scavenging assay asymtotes?

Hello. I am here looking for help with the NO radical scavenging assay. I was working on setting up the assay with sodium nitroprusside and the Griess reagent as they do in most works. However, I...

27 May 2024 5,013 1 View

Inconsistente results with cell line LX-2, any advice?

Hi there! I'm working on my master's thesis and for my project I'm testing a new curcumin derivative in liver fibrosis. I'm working with the cell line LX-2 and have been having problems since the...

21 May 2024 2,171 0 View

Will ultracentrifugation be replaced as the most widely used method for isolating extracellular vesicles?

Ultracentrifugation is the most widely used method for isolating extracellular vesicles (EVs), despite the significant structural damage that EVs suffer during the process.

16 May 2024 9,432 1 View

¿What control can I use to inhibit the proinflammatory effect of LPS in macrophagues?

I want to evaluate the polarization to M2 phenotype in Raw Cells, I co-cultured the cell with LPS and my drug to see the effect of the drug to inhibit the proinflammatory effect of LPS, but I have...

05 May 2024 8,525 0 View

Scientific support for research in Dentistry University of Los Andes Venezuela?

Sirs. ResearchGate Receive a cordial and respectful greeting. For academic purposes only, we request collaboratively the following articles for one of our research professors, who is preparing...

05 May 2024 9,459 9 View

How can we prevent small peptides (1000 Da) from eluting out of the gel during staining in a peptide gel (tricine-SDS-PAGE, Shagger)?

How can we prevent small peptides (1000 Da) from eluting out of the gel during staining in a peptide gel (tricine-SDS-PAGE, Shagger)? There are different types of fixing solutions, but I'm not...

28 April 2024 1,522 0 View

Which are Australia regulation for disposal of produced water from oil and gas activities?

I am currently conducting research for my PhD on environmental regulations, specifically focusing on the disposal of produced water from oil and gas activities in Australia. I would appreciate any...

18 April 2024 1,544 1 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

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 View

Torsten Wurm

In the past I tried to transfect HUVEC cells and the short answer is , they are very hard to transfect with Lipofectamine or similiar reagents. Lipofectamine LTX worked the best if used with OptiMem to dilute the DNA. The best method which worked for me however was to use an adenoviral vector.

Marcel HJ Ruiters

We used SAINT-MIX for HUVEC as is published years ago. The publication is attached. You can found more and recent info at the Synvolux site.