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Pippuri
Member since: Jul 12, 2009
From: Georgia, United States
Status: Protein Chemistry & Glycobiology Moderator
My points: 1063    what's this
Name: [Privacy]
 


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Feb 12, 2015    07:28 PM

Replied to topic in Cell Culture and Tissue Culture forum   Cell loss during annexin staining/flow cytometry

Hi Sophie, what is the source of neutrophil you use in your experiment? Did you observe any clumps while handling them?

Feb 5, 2015    08:12 PM

Replied to topic in Cell Culture and Tissue Culture forum   does MG132 dissolve in culture media? pls im desperate for help

Hi Annafreud, There are couple possibilities- How did you prepare MG132? While MG132 is very stable in organic solvent, it is rather unstable in aqueous condition. It has to be prepare fresh in the aqueous solution (ie. media) on the same day of experiment. The cells that you are treating M ...

Feb 5, 2015    05:43 PM

Replied to topic in Cell Culture and Tissue Culture forum   does MG132 dissolve in culture media? pls im desperate for help

Hi Annafreud, It isn't clear to me the problem that you are encountering is either the irreproducible results or the cells are dying (or not dying). Can you please elaborate?

Feb 4, 2015    10:47 PM

Replied to topic in Biochemistry forum   Sequence specific pulldown of dsDNA

Hi Baconman, In theory, it should work. I guess it is just a matter of optimization, which requires patience of a saint! :-) Good luck.

Nov 26, 2014    08:06 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Detecting small MW protein via SDS-page / Western

Loulou34, How much methanol or ethanol you have in your transfer buffer?

Oct 20, 2014    05:26 PM

Replied to topic in Neuroscience forum   PC12 and voltage gated calcium channels

Hi Domino, You can use ω-Conotoxin MVIIC for blocking N and P/Q type Cav channel. Good luck.

Aug 28, 2014    04:36 PM

Replied to topic in Cancer forum   Western blot analysis

Hi Britjade91, What is the blot corresponding to the imageJ histogram? phospho-ERK? Does the blot give you one or two bands? Depends on the source of your sample, some time you see one band, some time you see two... If you are getting two bands and both of them are your target, then I would gr ...

Aug 21, 2014    04:23 PM

Replied to topic in Grants, Manuscripts, and Career Advice forum   post doc salary and living expenses - usa

Hi gsitara11, Many PIs pay first year post-doc per NIH guideline. Right now it is USD 39264 per year. But the number is negotiatable- some pay more than that, some pay less than that. Living expenses really depend on which part of the country you are looking are. Even in the same state, living ex ...

Jul 21, 2014    09:54 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Problem with SDS-PAGE

Hi kola, One possible explanation leads to such staining pattern is bacterial growth in the running buffer. Try to prepare fresh running buffer from scratch and store it in a fresh container and run a gel again. Good luck

Jul 18, 2014    07:08 PM

Replied to topic in Biochemistry forum   western blotting

Hi milsey, Try to incubate your sample at 56 to 70 degree for 15 min instead of 95 to 100 degree often help with keeping membrane proteins in solution and may help with your detection. Good luck

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