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Questions related from Antonio Rosato
Hi to all I am writing you in order to get more information about the procedure 3.2.1 on Arsenic determination in glass containers for pharmaceutical use. In the monograph, at the paragraph...
06 June 2018 1,337 3 View
hi to all, i'm experiencing several issues with detection of elements such as sulfur and osmium at icp-oes. in particular, i work with an Agilent ICP, and at each analysis, i have to analyse these...
09 September 2017 5,461 3 View
hi to all, i would like to estimate the superficial pH on spherical particles having dimension of nanometers. since the surface is regular and omogeneusly coated, i was wandering if could i infere...
03 March 2016 8,999 2 View
hi to all, I need to use amicon ultra filtration devices at low Temperatures in order to purify a protein sensitive to this parameter. can i use amicon devices in a centrifugue refrigerated at 4°C?
03 March 2016 8,269 4 View
hi to all, i was wandering if it is possible to transform bl21 de3 cells with two plasmids. the aim is to espress 2 proteins at once. the two plasmids are both iptg inducible, and ampicillin...
02 February 2016 3,067 8 View
hi to all, i was wandering if it is enough to pellet cells (e.coli) from the protein induction medium containing iptg and resuspend them in a new medium without inductor to stop the protein...
02 February 2016 3,602 3 View
If I add chloramphenicol only in the overnight liquid culture, is ok? should the chloramphenicol be present also during the induction of the protein? thank you
01 January 2016 1,215 3 View
hi to all, i have noticed that if i incubate two parallel bacterial cultures in the same conditions ( one where i add IPTG for protein induction and another in which I don't add anything) the OD...
01 January 2016 1,020 7 View
Hi to all, I'm wondering if glucose polymers such cyclodextrins, glycogen and more complex polymers, are metabolized by E.Coli. Thank you
10 October 2015 2,756 4 View
hi to all, I'm trying to dock a 4Fe-4S cluster into a protein using the Haddock web-tool. I suppose that the protein I'm using coordinates the cluster by means of specific residues, that I have...
06 June 2015 5,931 0 View
Hi to all, I'm trying to express and purify a protein. Once the lysate was obtained, I performed an ion exchange chromatography followed by a size exclusion. In the chromatogram of the SEC, i have...
05 May 2015 3,511 9 View
Hi to all, I have to dock a protein with a protein. I have determined the active residues by means of CSP. I know that when the two proteins interact, share a copper ion, that is that is a sort of...
03 March 2015 9,480 1 View
Hi to all, I have to analyze an in-solution tryptic digest performed in the presence of 0.1% sds. I'm wandering if I can directly analyze it by means of rphplc ms w/o desalt it with zip tip since...
03 March 2015 3,131 3 View
Hi to all, I'm trying to analyze by mass spec. a DNA Binding protein having a particular conformation, so I'm interested in isolate it and analyze it as intact by mass spectrometry. I've tried to...
03 March 2015 8,561 11 View
Hi to all, I'm trying to study a protein DNA complex. when I try EMSA i have positive results with the nuclear extracts, while with the recombinant protein I have never seen any interaction. On...
03 March 2015 5,281 6 View
Hi to all, I have to perform an experiment in order to evaluate the behaviour of a protein as the dielectric constant of the bulk varies. So I thought to study the extrinsic fluorescence of the...
02 February 2015 6,133 2 View
Hi to all, I'm approaching to the haddock web-tool for the first time. I got the username and password for the easy interface. I'd like to know wheather i'm on the right way. Once I've uploaded...
12 December 2014 7,044 5 View
Hi to all, I'have to perform a docking in haddock. i have determined the most susceptible residues upon ligand binding by chemical shift perturbation. now i need to determine which among the are...
12 December 2014 7,742 8 View
Hi folks, I'd like to know how can I detect whether a protein has been incorporated into a liposome, and how much of it is entered the vesicle. What kind of experiment can i perform? thank you...
11 November 2014 5,650 8 View
hi to all. Is it possible to remove the blocked milk proteins from a pvdf membrane?
09 September 2014 5,566 2 View
Hi to all. is it possible to analyze by MS spec a band excised from a milk blocked PVDF membrane? could i have any problems dealing with the use of milk?
09 September 2014 6,851 3 View
hi to all, is SDS compatible with zip tip? i need to eliminate sds from a protein sample containing 10% of this detergent, can i use ziptip? otherwise could i use alternative detergents such as...
09 September 2014 9,103 5 View
Hi to all, I have expressed and purified a 6x his tagged metallothioein like cysteine rich protein. By means of various techniques such as electrophoresis and NMR, I have realized that the protein...
09 September 2014 1,544 4 View
Hi to all, AMBER and CHARMm are among the force fields used for computate protein properties. I'd like to know Which are the differences between the AMBER and the CHARMm Force fields, and when...
08 August 2014 6,191 4 View
Hi to all, As in object i'like to know the amino acid sequence of the NLS of a Nesprin-1 protein. Thank you
08 August 2014 7,342 2 View
Hi to all, I'd like to perform an immunoblotting against a protein on my EMSA. I would like to know if I can strip the streptavidin-HRP / biotinylated DNA probe complex from the membrane and...
07 July 2014 1,654 2 View
hi to all, I need to quantify a metal in order to understand if it triggers the binding of a protein to DNA. So I performed an EMSA and I transferred the gel on a 6,6 Nylon membrane positively...
07 July 2014 3,202 15 View
Good morning, I need to analyze trace metals (such as Copper) in biological samples. Specifically I need to determine the distribution of the same metal in the cytosol and in the nucleus after a...
07 July 2014 674 12 View
Hi to all, The cross linking step in the EMSA assay is performed to covalently bind the DNA probe to the positively charged membrane. Does the protein DNA interaction turn covalent too due to...
07 July 2014 2,293 1 View
Hi to all, do you know any database where you can submit the peaks of ESI mass spectrum of an entire protein with its multi charge states, and the database can match that spectrum with a protein?
07 July 2014 399 16 View
Hi to all, Is there any method mass-spec compatible to extract proteins from an agarose gel and purify them so that I can identify them by a direct injection in a ESI-MS? For example if I...
07 July 2014 6,695 7 View
I'm wondering whether an interaction between copper I ions and DNA is possible and to what extend this interaction could affect the DNA conformation. Do you know any literature talking about this...
05 May 2014 1,224 3 View
I have electroblotted on a pvdf membrane a 4-20 PAA-TGS gel run with samples 3 lanes containing DNA and a recombinant protein and 3 lanes of control containing only the protein. All the sample...
04 April 2014 9,686 14 View
I've performed a week ago a western blot in a membrane, and once I completed my assay, I conserved the membrane in my notebook. Now I'd like to perform some assays, for example with mass...
04 April 2014 5,123 4 View
In my last experiment was happened such a puzzling thing: I performed an electrophoresis (using a 4-20% TGS PAA gel) on samples containing Both a ~30 bp DNA duplex and a recombinant protein...
04 April 2014 5,679 8 View
I've performed an EMSA assay. I have run a biotinylated free probe and my samples with the protein. In all the lanes I observe shifts, also in the control lane. I suspect that in the control lane...
04 April 2014 5,319 2 View
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03 March 2014 7,687 3 View
I am wondering if it is possible to perform a western blot on a gel after it has been destained from a silver-staining.
03 March 2014 8,163 2 View
I'd like to know if anyone has ever used the anti-atox1 antibody and above all if anyone knows its molecular weight?
03 March 2014 6,500 2 View
The role of ionic force on the protein-DNA complex's formation is well known. There are some EMSA protocols in which the binding buffer is prepared with NaCl and KCl, others in which there is...
03 March 2014 5,650 0 View
In some protocols it is advised to incubate the protein and the DNA in the binding buffer on ice, other protocols advise to perform the binding reaction at room temperature. How to manage this...
03 March 2014 4,696 5 View
I'm just wondering if this kind of staining is linear with the concentration of DNA.
03 March 2014 3,912 1 View
