Why shRNA control cell line behave like that ?

I have KO cancer cell line, and I am applying shRNA to target some genes. My control is RFP-shRNA. I ran western blot to check my protein of interest, and i have found that KO cell line with...

08 September 2018 5,117 6 View

Why flow cytometry and Qpcr are different ?

Hello everyone, I have unexplained phenomenon in my data regarding one cytokine. The level of this cytokine by Qpcr is very high in knockout more than WT mice in naive condition and after...

08 September 2018 4,417 3 View

How long should I stimulate cell line for NGS ?

Dear fellow, I am planning to send samples of cancer cell lines to NGS. I have one set unstimulated, and one set with cytokine stimulated. However, I am not sure if it is better to stimulate...

05 June 2018 9,997 2 View

What is this bacteria ? how to get rid of it ?

Dear bacteria and cell culture specialists, Recently we encountered this bacteria during cell culture, please see the attached images. These bacteria doesn't change the PH of the culture media...

03 April 2018 2,510 4 View

Can I use sonication in EMSA?

Hello, I'm doing EMSA using adherent cell line . I extracted the nucleus part but it was very viscous. Why the nuclear extract was very viscous ? And can I apply sonication ? Thank you.

03 April 2018 8,852 4 View

Why DNA migrates slower than DNA/protein complex ?

Hello fellows, I am doing EMSA. I wanted to check for the binding and efficacy of DNA and nuclear extract proteins. So, I run Acrylamide gel (native, 5% , 0.5xTBE buffer, with DNA view dye )....

03 April 2018 7,945 23 View

How to find a promoter sequence for specific gene ?

Hello fellows, How can I find a promoter sequence for a certain gene ? ( simple way to explain , i don't have much bioinformatic background) Thanks in advance

02 March 2018 9,567 11 View

Double band in western blot ?

Hello guys, I am running western for the 3rd time for the same protein. First two times I can see clear one band. It is inducible protein. However, on the 3rd time, I have two bands , instead...

01 February 2018 2,616 2 View

Flag-tag on C-terminal or N-terminal?

Hello Fellows, I want to clone a protein with a Flag but I can't decide where should I place the Tag on C-terminal or N-terminal since some papers put it on N-terminal but others put on...

10 November 2017 1,280 4 View

How to measure Calcium ?

07 August 2017 948 6 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 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

Compact antenna for the frequency range 1 MHz-5 MHz?

Hello everybody! I have a problem with developing a compact antenna for the 1MHz-5MHz frequency range. Tell me what type of antenna can be used and what books and articles to read to perform this...

27 February 2021 7,710 4 View

Why do the air flow results vary totally when adding scalar in CFD ?

Dear All, We are using Autodesk cfd to calculate the air flow in a single sided naturally ventilated room, where air flow depends only on buoyancy due to temperature difference between inside and...

25 February 2021 9,602 4 View

No amplification data for qPCR on MxPro?

I have set up a qPCR run on MxPro with SYBR green and ROX as a reference, however, when I go to the analysis section the program seems to only collect data for the dissociation curves and not the...

25 February 2021 8,039 1 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

24 February 2021 8,582 1 View

Does PHA-P stimulate the proliferation of lymphocytes from mice?

Hello, I will start performing MLR using lymphocytes from mice's spleen and I would like to know the better way for the stimulation. If is using PHA-P or CD3/CD28? I would like to know as well if...

23 February 2021 5,476 4 View

Why anti-CD20 or other antibodies are not used to label target cells in flow cytometry t-cell cytotoxicity assay?

New to the field of cell therapy. Why fluorescent-labeled antibodies to target cell antigens are not used for determining target cell count by flow cytometry in cytotoxicity assay (eg car-t)? Cell...

21 February 2021 3,278 3 View

How to calculate time-averaged fields from Ansys CFX solver?

I would appreciate any comments on the following problem. I need to calculate mean fields (e.g. pressure, velocity and vorticity), for which I activate Trn Stats tab in CFX-Pre. However, what I...

21 February 2021 8,548 7 View