Is there a "good" transDUCTION protocol?

Hello dear colleagues, I am experiencing strange problems when performing transDUCTION on my CD4/CD8 cells (separated using Miltenyi beads from total PBMCs from a buffy coat). For now on it is...

03 April 2016 6,843 1 View

Any limitations for expression in T-lymphocytes?

Hello everyone, we are trying to transduce T-lymphocytes (TLs) with vector containing IRES, and cannot see any expression using flow cytometry (our target protein is conjugated with GFP). The...

02 March 2016 6,562 4 View

Is it possible to overcome co-expression?

Hello, in my expression cassette I already have 2 proteins that are expressed using IRES. However I'd like to express another protein at the same time (a homing molecule), but, honestly, I do not...

01 February 2016 5,983 3 View

A good biotin conjugate for FACS?

01 February 2016 8,243 5 View

What's the best solution for T cells expansion?

Hi, I am trying to perform an expansion of T-cells, purified (by miltenyi columns) from PBMCs and transduced with construct of interest. I am wondering if there's a reliable and "working" protocol...

11 December 2015 386 5 View

How avidin interacts with steptavidin-binding protein?

06 July 2015 5,957 0 View

Best lab journal software?

Hello, everyone. We all know how, sometimes, it's difficult to write down everything into a lab journal, while I believe there are other approaches apart MS Word :) Would anyone be able to share...

06 July 2015 5,357 0 View

What's the "normal" percentage range for PAN-T cells out of PBMCs?

Hi, colleagues. Unfortunately I have just started to work in this new field for me, PBMC purification from blood bags, MACS separation etc. So what I do is out of a blood bag from hospital I do a...

05 June 2015 5,812 8 View

Do I need to preculture cell for siRNA experiment?

Hello, I am planning to perform a siRNA knockdown using lifetechnologies Lipofectamine RNAiMAX reagent, so taking into account they require 60-80% confluence of cells on the moment of transfection...

02 March 2015 1,424 5 View

FACS markers for Jurkat analysis?

Hello, I am about to complete my FACS training so I need to bring some cells labeled with different markers (correct me if I'm wrong in terms). What I have here in culture are Jurkat cells, so I...

01 February 2015 7,338 3 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 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