Dear all, i would like to conduct some immunofluorescence staining on cell lines, but i am clueless about the things required for the assay?

More Justin Tan's questions See All

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 managed to purchase a Ki67 antibody with a conjugated fluorescent marker. How do i use this to quantify cell proliferation in 3D culture?

As above. Thank you :)

10 November 2017 6,237 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

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Axel Heiser

I recommend reading:

Fluorescence Microscopy: A Concise Guide to Current Imaging Methods, Christian A. Combs. DOI: 10.1002/0471142301.ns0201s50

Also if you google "fluorescence microscopy protocol" several good protocols come up.

Good luck,

Axel

Justin Tan

Dear Sir,

Thank you for your suggestion. Greatly appreciated.

Warmest regards,

Justin

Dear sir, I am fine with staining dead cells. The cell lines are adherent.

For your kind advice, please.

Thank you.