Best recipe for Ripa buffer?

Hello, today i want to ask about ripa buffer, buffer used in Western blot.(Very simple but seemingly important question) You may already know this, but i found out that there are numerous...

09 October 2017 6,617 3 View

phospho form and total form proteins in Western blot?

Hello, I want to ask about the method of analyzing phospho form and total form of proteins by western blot. I learned that many people conduct stripping the membranes to study expression level...

09 October 2017 2,290 2 View

Research on protein whose localization depends on cell density

06 July 2017 6,986 6 View

Cautions during detection(Western blot)?

Hello, Today i want to ask about films used for detection during western blot. When i conduct detection, i know quite well that film should not be in developer solution too long to avoid...

03 April 2017 6,239 3 View

Phospho form and total form antibody?

Hello Today i want to ask very simple but seemingly important question. When conducting western blot, i have to see the expression of phospho form of protein(Protein of my interest). To measure...

01 February 2017 2,790 3 View

Quetions or comments about results of western blot?

Hello, I always get great support and help from Researchgate, And i am sorry to anyone whom i forgot to express my gratitude for reply. Some days ago, i asked same question about sample buffer...

10 November 2016 4,492 4 View

Method for extracting proteins for western blot?

Hello, Today I want to ask whether there is absolute way of extracting proteins from cells for western blot. I found out that there are numerous ways to extract proteins. In my case, I treat...

09 October 2016 6,334 7 View

Dilution rate of developer and fixer in western blot?

08 September 2016 9,924 3 View

Questions about developer and fixer?

Hello, I am always grateful for those who answer my questions. I learn a lot thanks to such help. I want to ask question about developer and fixer, required for western blot. I asked quite a...

08 September 2016 3,344 2 View

Important or unimportant factors during western blot?

07 August 2016 7,991 6 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

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

No title

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

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