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

I need a straight protocol to study deposited ECM in cultured cells upon stimuli with selected molecules

08 September 2019 836 2 View

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 891 11 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 9,254 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 6,353 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 9,786 25 View

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

01 January 1970 9,750 6 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

23 February 2021 4,014 3 View

How to reduce static electricity of powder after lyophilization?

I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...

22 February 2021 10,099 5 View

Can we remove the fmoc group after peptide cleavage?

I have synthesized the peptide using Fmoc based solid phase peptide synthesis. Also I have cleaved the peptide from the resin and lyophilized it, but later realized that, I forgot to deprotect the...

21 February 2021 4,537 3 View

How T cells recognise non-peptide antigens?

This might sound very basic question, but please share with me an explanation of how the T cell is activated when the antigen is not a peptide? What happens? And what are the molecules involved in...

21 February 2021 564 3 View

Help with Amyloid beta detection protocol (western-blotting)?

Hi everyone, I am a first year PhD student working on amyloidosis. I would like some tips and tricks to crack efficiently my current nightmare, i.e. the western blotting of the Amyloid beta and...

11 February 2021 2,513 3 View

How to assing secondary structure to calculated peptide/protein having xyz file?

Hi, I have an optimized structure of some peptides and I would like to represent it as a secondary structure - cartoon. Ok, so server apps ask for pdb file (like Stride). I tried in different...

04 February 2021 517 3 View

Identical spectra for L- and D-peptides after they self-assemble?

Hi dear all, I need advice. I have two peptides with the same sequence but one consists of L-amino acids only and the other consists of D-amino acids only. when they self-assemble in aqueous...

03 February 2021 2,032 3 View

FLAG purification, elution doesn't have protein?

I am purifying a heterotetramer, with one subunit having a 1D4 tag and the other having a FLAG tag. I purified from Sf9 with HBS pH7.3 and 0.002%LMNG first with Anti-1D4 resin and got an adequate...

02 February 2021 3,860 1 View

How long can synthesized peptides using SPPS be stored? Storage Conditions?

I have been synthesizing peptides on resin beads. I am wondering what best storage method is there to store the beads after synthesis before cleaving. And how long can they be stored in that...

02 February 2021 5,636 5 View