Best cell lysis solution for DNA extraction by manual method?

Could anyone help me find the best composition for cell lysis solution for manual DNA extraction using blood as the source? I want to prepare the solution in house. Does anyone have experience...

10 November 2018 2,913 5 View

How to ascertain/check the quality of MilliQ or ELGA water?

We have an in-house ELGA water system (something like MilliQ water system). We need to check its quality? Can we do it in-house? If so, how?

02 March 2016 3,582 3 View

Compensation in flow cytometry

Can I compensate with beads for my experiments. I use 4 colours in a single tube (FITC, PE, PerCP Cy5.5, APC). What does it matter if I use beads for my compensation. Why can't I just use beads to...

02 March 2013 5,259 4 View

Trypsin edta

Would anybody know if there is any difference in using DI-Sodium and TETRA-Sodium EDTA in the preparation of 0.25% Trypsin - 0.53mM EDTA? Thanks!

31 December 2011 2,575 0 View

Good protocol for ICC/IF for cultured cells

Can anyone please lead me in getting a Good protocol for ICC/IF for cultured as well as PFA fixed cells? I am working on oral tumour tissues and cell culture. Thanks for your time.

31 December 2011 3,115 5 View

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 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

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28 February 2021 9,936 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View