What is the correct manner to represent the effects of a treatment in autophagic flux assay in vitro?

Dear all, In an in vitro experiment of autophagic flux assay by western blot using chloroquine, is it correct to use the LC3-II/LC3-I ratio as a manner to represent the effects of a treatment or...

07 August 2018 5,297 0 View

Does anyone know if 25 or 50 uM of chloroquine treatment could be toxic for the cells (C2C12 cells or any other cell line) in a time-course treatment?

Does anyone know if 25 or 50 uM of chloroquine treatment could be toxic for the cells (C2C12 cells or any other cell line) in a time-course treatment like 1, 3, 24, and 48 h?

05 June 2018 117 1 View

Should I add 0,1% BSA to the serum-free media to produce the C26-conditioned media or just culture the cells in the serum-free media is sufficient?

Hi everyone, I want to produce conditioned media from C26 cells for then treat C2C12 myotubes with this conditioned media. I have read some papers using serum-free media for 24h to produce C26...

04 May 2018 1,589 0 View

Should I add 0,1% BSA to the serum-free media to produce the C26-conditioned media or just culture the cells in the serum-free media is sufficient?

Hi everyone, I want to produce conditioned media from C26 cells for then treat C2C12 myotubes with this conditioned media. I have read some papers using serum-free media for 24h to produce C26...

04 May 2018 7,667 0 View

How would you all interpret the following results of autophagic flux?

Dear all, How would you all interpret the following results? Please note the cells were incubated for 24 or 48h in conditioned medium in the presence of chloroquine during whole time-course of...

01 January 1970 6,310 4 View

What is the correct manner to represent the effects of a treatment in autophagic flux assay in vitro?

Dear all, In an in vitro experiment of autophagic flux assay by western blot using chloroquine, is it correct to use the LC3-II/LC3-I ratio as a manner to represent the effects of a treatment or...

01 January 1970 981 3 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

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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