What is the method or buffer conditions for lysing mammalian cells for both genomic AND mitochondrial DNA?

Hi all, I am wondering as to the conditions of a lysis buffer that would lyse both the cell membrane and the mitochondrial membrane. The purpose of this lysis is to collect genomic and...

09 October 2014 418 3 View

Bacterial whole cell lysis?

I'm looking for a lysis buffer recipe and protocol for lysing an E. coli strain. The background is that I will be using IPTG to induce the expression of a plasmid with a specific cDNA. I...

10 November 2013 6,167 6 View

Techniques to destain Giemsa-stained cell colonies

I am currently staining cells to quickly look at the effect of a drug for both 24 hour and continuous treatment. At the end of the assay, the cells are to be stained with Giemsa and will be judged...

31 December 2012 2,909 8 View

Is the ChIP assay the only verified assay for chromatin-bound proteins?

Currently wanting to look at chromatin-bound vs soluble protein concentrations.

09 October 2012 9,952 0 View

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02 March 2021 3,060 3 View

Where can I find life cycle inventory or assessment of non woven materials?

Dear network, I am looking for the LCA or LCI or at least the energy balances of the non woven fabrics process for my master project, in specific for natural fibers, yet I have found...

02 March 2021 9,662 1 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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 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

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28 February 2021 9,936 2 View