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Member since: Jul 12, 2009
From: Georgia, United States
Status: Protein Chemistry & Glycobiology Moderator
My points: 1055    what's this
Name: [Privacy]

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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

Jul 17, 2014    12:11 AM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   yield of purification by IP and Co-IP

Hi dorvashmr, One option, which can definitely get you a lot more proteins than IP is to get a plasmid that carry your protein of interest, overexpress it in E. coli and perform a quick affinity purification. It may take longer time (having to do transformation, grow the bugs and finally purific ...

Jul 12, 2014    12:36 AM

Replied to topic in Biochemistry forum   WESTERN BLOT - high molecular weight protein

Hi ndora, If you only loaded 100ng of cortical and hippocampal lysates per lane, I suggest you to go higher amount than that. Says, 20 to 30ug. Also, for transferring a protein that large, you may want to transfer for longer time- 20 to 30 V for 16 hours at 4 degree. Another thing is to dro ...

Jun 12, 2014    04:26 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Protein-oligonucleotide gels

Hi Mi: Yes, if you perform the electrophoresis on a 20% SDS-PAGE. Good luck

May 13, 2014    04:25 PM

Replied to topic in Cell Culture and Tissue Culture forum   determining GLUT4 to the cell membrane bound

Hi lunaoreaD, For muscle cells and adipocytes, both of which express GLUT4 endogenously, you do not need to overexpress the transporter, given that you have the proper antibody to detect the endogenous form of GLUT4. Good luck.

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