Search found 13 matches

by claire84
Tue Jan 15, 2013 11:12 am
Forum: Bioinformatics
Topic: scatter plot program
Replies: 1
Views: 10770

scatter plot program

Hi everybody!
Can you recommend me a simple and free program to use to perform 3D scatter plot?!
Thank you
by claire84
Fri Nov 02, 2012 2:56 pm
Forum: Cell Biology
Topic: peripheral mononuclear cells handling
Replies: 3
Views: 4057

Re: peripheral mononuclear cells handling

Indeed want to study monocytes. I think I'm going to use PBS/edta and to scrap them with the tips of the pipette.
Do you think it's a good idea?
by claire84
Thu Nov 01, 2012 12:18 pm
Forum: Cell Biology
Topic: peripheral mononuclear cells handling
Replies: 3
Views: 4057

peripheral mononuclear cells handling

Hi guys, maybe it's a silly question, but I'm not a cellular biologist and I need some help. I isolated PBMC from the whole blood and then I seeded them in a 96 well plate. After incubation of 24h I would like to monitor the presence of a certain protein on their surface using a labelled antibody. D...
by claire84
Wed Sep 05, 2012 4:15 pm
Forum: Cell Biology
Topic: HL60 cell line suppliers
Replies: 0
Views: 2313

HL60 cell line suppliers

Hi,
anyone knows where I can buy HL60 cells with a low passage number?
thank you so much
Claire
by claire84
Wed Jul 04, 2012 3:58 pm
Forum: Cell Biology
Topic: HL60 cells centrifugation
Replies: 1
Views: 2666

HL60 cells centrifugation

Hi everyone! I need your help because I have little experience on cell culture techniques...I would centrifuge my Hl60 cells which are plated in a 96-well microplate, to obtain a pellet at the bottom and to remove supernatant without loss of cells. Could you recommend me rpm and time to use? I tryed...
by claire84
Fri Jun 15, 2012 2:05 pm
Forum: Cell Biology
Topic: HL60 cell line shedding
Replies: 0
Views: 1768

HL60 cell line shedding

Hi guys!
I need your help!
I have a HL60 culture and I'm searching for something (chemicals or interleukins or other factors) that can cause the shedding of PSLG-1 protein from the cell surface. Can you hel me?
thank you in advance!
by claire84
Thu Jan 12, 2012 11:15 am
Forum: Molecular Biology
Topic: enzyme inhibition assay
Replies: 0
Views: 2145

enzyme inhibition assay

Hello everyone, I need some help to interpret my results...I'm working with anthrax lethal factor, which is a Zn2+ dependent metalloproteinase. I would like to calculate Ki of an inhibitor using a FRET inhibition assay.The FRET assay are run for 30 minutes at 25 °C. At the end I obtain a graph inten...
by claire84
Tue Jan 03, 2012 4:26 pm
Forum: Cell Biology
Topic: His-tagged proteins
Replies: 2
Views: 3250

Re: His-tagged proteins

In our lab, we do not produce recombinant protein, we purchase them for inhibition assay....
but I think I could try to use his-tagged hiv-protease, N and C termini are not closer to active site

thank you for your reply!!!
by claire84
Tue Jan 03, 2012 9:13 am
Forum: Cell Biology
Topic: His-tagged proteins
Replies: 2
Views: 3250

His-tagged proteins

Hi everybody, I have some doubts about the use of his-tagged HIV-1 proteinase for an inhibition assay. I have to test some potential inhibitors and I would like to perform a kinetic assay to obtain the percentage of inhbition and the IC50 values. Can the presence of the tag influence my results? Is ...
by claire84
Tue Oct 11, 2011 2:47 pm
Forum: Bioinformatics
Topic: Pairwise local alignment
Replies: 3
Views: 4672

Re: Pairwise local alignment

Yes, I'm looking for similar residues in the active site of tese two proteins. So, I have to perform cycles of local alignment using different substitution matrix. Is it right? Can I use LALIGN?

thank you!
by claire84
Mon Oct 10, 2011 4:32 pm
Forum: Bioinformatics
Topic: Pairwise local alignment
Replies: 3
Views: 4672

Pairwise local alignment

Hi,
I'm not a bioinformatic, but I need to perform a sequence alignment between two protein which are not very similar. In this case is better to perform a local alignment, right?
I did this kind of aligment with LALIGN tool.What do you think about this tool?

THANK YOU IN ADVANCE!
by claire84
Mon Aug 23, 2010 3:41 pm
Forum: Molecular Biology
Topic: enzymatic inhibition assay
Replies: 2
Views: 3696

I mix it in an eppendorf by means of a micropipette before to start the measure.The inhibitor is a reversible one. In the reaction mixture there are only the enzyme, the substrate and the inhibitor dissolved in the buffer described before