Can someone provide a protocol to study Extracellular matrix in cultured cells by proteomics?

More Alberto Vitali's questions See All

Can anybody suggest mixtures of phospholipids to better mimic bacterial and mammals membranes?

I need to test some peptides by using vesicle models

09 October 2015 1,124 11 View

Does anyone knows the mechanism of action of the PGRMC1 inhibitor AG-205?

I need this information to discriminate the action of AG-205 from that of a peptide

09 October 2015 3,038 1 View

My ZR-75-1 breast cancer cell line grows very very slowly, does anyone have experience with this cell line?

I bought this cell line from ECACC as a frozen sample, and three weeks after thawing its growth is almost undetectable. I'm using the medium recipe recommended.

02 March 2013 9,377 19 View

Is anyone using the 96multiwell microplates GSH coated for immobilizing proteins and detect peptide-protein interactions? Which protocol is the best??

I'm tryng to detect peptide-SH3 domain interaction by using the Thermo GSH coated microplates, but our results are not consistent. Perhaps the buffers we are using are not the best? And how can I...

31 December 2012 6,494 0 View

Same protein found in different spots (different pIs and MWs) in a 2D gel, isoforms?

When I ran a sample of Candida lysate in a 2D gel, I found the same protein in different points of my gel. After LTQ-Orbitrap MS/MS analysis it results the same proteins, and the tryptic fragments...

05 June 2012 9,900 25 View

Does anyone know a method/protocol to allow bacterial ribosome subunits to ri-associate?

I am purifying 30s and 50s ribosome subunits to successively perform studies on their ri-association upon inhibitor addition.

01 January 1970 9,859 6 View

How to define an anisotropic material with asymmetric elastic compliance/stiffness matrix in ANSYS APDL?

I need to model an anisotropic material in which the Poisson's ratio ν_12 ≠ ν_21 and so on. Therefore, the elastic compliance matrix wouldn't be a symmetric one. In ANSYS APDL, for TB,ANEL...

09 August 2024 5,048 2 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is it true that $\det(V(A))$ may be only $\pm 1$, depending on $n$, for the last symmetric tridiagonal matrix $A$?

One can try to generalize the Vandermonde determinant in the following direction: Let $A$ be any symmetric $n$-order square matrix. Consider its powers' diagonal elements $(A^k)_{ii}$ and...

08 August 2024 6,690 1 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Usage of internal standards in LC-MS/MS analysis?

Have you ever seen a LC-MS/MS method uses both internal standards and external standards (in matrix matching purpose) but the concentrations of internal standards are outside the calibration curve...

05 August 2024 3,084 6 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to use Density Functional Theory to calculate carrier mobilities of solid system?

Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...

04 August 2024 8,894 1 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

Alexandre Bruni-Cardoso

Have a look at the methods of this article by Ghajar et al Article The perivascular niche regulates breast tumor dormancy

Leandro Toshio Kochi

Hi Alberto!

Maybe this paper can help you with methodology and for reference:

http://essays.biochemistry.org/content/early/2019/08/26/EBC20190001.long