What is a good reference on designing experiments to determine molecular mechanism of disease or biological process?

I need to learn techniques and strategies in planning experiments that try to figure out the molecular mechanisms. Thanks.

01 January 2021 2,525 3 View

Has anybody had staining problems with lysotracker in live cells?

I am working with two fish cells types, a zebrafish liver cell line and primary cultures of trout macrophages. We are using lysotracker to stain them in order to visualize lysosomes in confocal...

16 December 2020 3,935 2 View

Is TRIS-EDTA-CITRATE buffer (pH 7.8) a good antigen retrieve for the TUNEL assay?

Hello everyone! I performed a TUNEL assay simultaneously with an immunofluorescence (SOX9 antibody) on the same sections of human testis. The fluorescence of TUNEL turned out well, but that of...

15 December 2020 1,270 3 View

Does a manager's background such as culture and attitude influence the performance of employees?

30 November 2020 1,585 2 View

If I want to use mean as calculation for my questionnaire, should I test the reliability and validity?

I'm using questionnaire for my research and mean (average) as my data analysis. I'm wondering how to calculate the reliability and validity of my data.

28 September 2020 3,517 3 View

How to induce with L-arabinose Chaperone Competent Cell BL21 (Takara)?

I am using Chaperone Competent Cell BL21 from Takara, pGro7/BL21 , but it is not clear on the protocol how and when I should induce the expression of the chaperones with L-arabinose. First time I...

14 September 2020 8,488 3 View

Is it better to use the arithmetic mean or geometric mean when averaging MPN?g or CFU/g measurements?

This data is from a microbiological evaluation of streetfood locally sold in our region. Thanks.

12 August 2020 5,981 3 View

Any recommendations on the software to do text mining or sentiment analysis on Chinese reviews?

I am trying to do text mining on Chinese reviews. I have tried out many softwares, like the RapidMiner, Chinese Text Analytics, Python. Most of them seem to require certain level of programming...

12 August 2020 9,109 8 View

Where can I find a good set of microbiological standards for aerobic plate count and coliform levels for street food or ready to eat food?

I need these to evaluate the data I obtained from my study. Thanks

10 August 2020 7,466 12 View

What is a good Thomson Reuters or Scopus indexed journal where I can publish my paper regarding Microbial safety aspects of street foods?

I need a journal with a faster turn around time, reasonable review process, and preferably no publication fees.

15 July 2020 8,241 3 View

No title

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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 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