Could I trust in qubit for protein quantification?

I need quantify the protein amout from human DCs lysed by RIPA buffer ( 150 mM NaCl, 1% NP-40 or Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, pH 8.0 ) to use in a immunoblot....

11 December 2019 3,701 3 View

Is it possible to use the same permeabilizing agent to perform intracellular staining of cytoplasmatic and nuclear proteins in flow cytometry?

I work with human mo-DCs and I need to detect cytoplasmatic and nuclear proteins by flow cytometry using flurochrome-conjugated antibodies (direct detection), that is, I need to permeabilize both...

04 May 2019 4,991 5 View

What is recommended temperature to perform fixation with formaldeihyde or paraformalde to intracellular staining by flow cytometry of culture cells??

04 May 2019 3,390 4 View

There is a need to fixation step to detect soluble proteins in cytoplasma or nucleus by flow cytometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, GBP1, GBP4) and nucleic proteins (IFI16), all apparently soluble, using direct detection by flow cytometry with...

04 May 2019 5,624 3 View

Why mercaptoethanol in THP-1 culture medium?

I have purchased THP-1 cell line from ATCC and they recommed mercaptoethanol. It is a reducing agent, what is the function of this kind of agent in culture medium of THP-1 cells? How...

04 May 2019 9,930 3 View

What is the best permeabilizing agent to detect soluble cytoplasmatic and nucleic proteins by flow cytometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, GBP1, GBP4) and some nucleic proteins (IFI16)using direct detection by flow cytometry with fluorochrome-conjugated...

04 May 2019 7,582 2 View

What is antibody concentration recommended to western blot?

I need to detect and measure the abundance of cytoplasmatic proteins (CASP1 an GSDMD) by western blot in human mo-DCs, and some companies where I purchased the antibodies do not specify a...

04 May 2019 1,385 5 View

What is the difference between formaldehyde and paraformaldehyde in cells fixation for intracellular staining using flow citometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, IFI16, GBP1, GBP4) using direct detection by flow cytometry with fluorochrome-conjugated antibodies in human mo-DCs and...

04 May 2019 6,491 4 View

How many human primary cells I need to quantify protein abundace in western blot analysis?

Someone that work with human primary cells and western blot: how many cells I need to identify protein abundance in western blot analysis? I want to quantify Caspase-1 and gasdermin D

31 December 2018 5,105 5 View

No title

I want to detect caspase 1 activation in human monocytes-derived dendritic cells after interaction with my pathogen.

10 November 2018 2,081 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

No title

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

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

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

Isolates of Hymenoscyphus fraxineus?

Dear Dr. Cai, my name is Simone Prospero and I work in the team of Phytopathology at the Swiss Federal Institute for Forest Snow and Landscape Research (WSL;...

01 March 2021 1,133 1 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