Is anyone has experience of transfecting hESC derived cardiomycytes? Which transfection reagent or methods can be used to transfect cardiomyocytes?

I would like to transfect human embryonic stem cell differentiated cardiomyocytes with GFP plasmid. Based on the literature, it does seems like, these cells are relatively difficult to transfect....

08 September 2016 6,029 4 View

Has anyone used Lipofectamine 3000 transfecting reagent to transfect H9 (human embryonic stem cells) cultured in mTeSR media?

I would like to use Lipofectamine 3000 (Life technologies) to transfect human embryonic stem cells (H9) and make the selection by sorting with FACS aria II. My basic concerns includes - Whether, I...

05 June 2016 2,443 2 View

What concentration of bleomycin one can use to induce ds DNA damage in HEK293 cells?

Further, how can you prepare concentrations of bleomycin? Can somebody suggest to me how exactly I can prepare bleomycin stock solution and what is the most efficient and appropriate...

01 February 2014 5,996 3 View

What is the correlation between DNA damage and the development of malignant tumors in cancer?

As DNA damages are mainly involved in the formation of primary tumor but somehow it diverts towards the development of melignant tumor, which leads to metastasis property. What is that mechanism...

31 December 2013 5,892 8 View

Is it possible to keep plates with fixed cells in half of the wells in PBS and live cell in other half with culture medium?

01 January 1970 5,210 1 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

No title

28 February 2021 9,936 2 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View