Is using NaOH for lysis of mammalian cells a good idea if you wish to extract a certain compound for LC-MS/MS?

Dear all, good day! I have a question regarding the use of NaOH for lysis of mammalian cells. Is it a common practice? Are there any publications which have used NaOH to perform cell lysis? If...

03 April 2019 5,332 4 View

Which genes or proteins involved in cell migration/motility should i probe for?

Dear all, recently i did a scratch test migration assay on my test compounds incubated with my cell line of interest and it appeared that the cell line seem to exhibit signs of increased cell...

10 November 2018 382 5 View

Should i use serum-free media for MTT assay?

Dear all, recently i am considering if i have been doing my MTT assay wrongly as i have been using my media (with 10% FBS) for my cell viability assay. I realised that the cells were dying at high...

10 November 2018 8,866 2 View

How do you carry out cell migration assay if you want to study if a drug has any effect of increasing cell migration towards another cell type?

Dear all, Recently i have a really crazy and brilliant idea to determine if a compound would be able to increase / decrease the motility of a cell type towards another cell type. How should i go...

09 October 2018 2,941 5 View

Cell death or drug precipitation??

Dear all, recently i found some strange and uncanny sights in the wells of cells treated with my compound of interest. Apparently when i checked the water solubility of the compound of interest,...

09 October 2018 1,427 3 View

Dear all, what are some of the assays i can use to show an increase in intercellular adhesions?

Dear all, i have recently discovered that one of my test compounds is able to promote intercellular aggregation (i.e. the cells are aggregating when treated with the compound). However, i can only...

04 May 2018 7,219 1 View

Dear all, do i require a sterile water control if i used sterile water as the solvent for my drug in cell viability studies?

Dear all, recently i started a cell viability study using presto-blue reagent for the testing. However, i dissolved the drug of interest in sterile water before adding into the wells (96 well...

04 May 2018 8,971 0 View

Dear all, i am currently working with HaCAT cell line. I wonder if you have any advice on the list of keratins which it can produce?

As above. Thank you :)

10 November 2017 9,256 1 View

Dear all, i would like to add gelatin and collagen as a liquid solution to my cell culture, but they are solids at room temp. Any suggestions?

As above. Thank you :)

10 November 2017 7,693 4 View

Gene and protein expression woes?

Dear all, Recently I ran qPCR for 3 different sets of treated samples and the trend of the mRNA seemed to exhibit an increasing trend with increase treatment for all the genes tested. However,...

10 November 2017 7,303 4 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

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02 March 2021 3,060 3 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 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

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 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

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 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