Dear Protocol-Online
I am running a series of native PAGE experiments, using Bio-RAD TGX 4-15% gels, and Tris-Glycine buffer pH=8.1
My protein of interest has a pI of about 6, and I am investigating it's interactions with some multimeric proteins.
When running the experiment I usually prepare lanes as follows:
1)multimer alone
2)multime + protein
3)protein
I use silverstain for visualization.
I do not observe a direct colocalizaion of my protein on the multimers, but it surely changes its
pattern, and the protein in lane 2) does not wonder as far as the protein alone in 3).
What can I conclude from an experiment like this?
How du you like my setup for native PAGE in general?
And yeah, this is my first time with native PAGE.
best regards
Peter Jallerup
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4 replies to this topic
#2
mdfenko
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Posted 27 January 2012 - 08:24 AM
a picture would have helped but...
if lane 2 is slower moving than and there is no band at the same place as lanes 1 and 3 then you are seeing interaction.
your experimental design appears to be okay
if lane 2 is slower moving than and there is no band at the same place as lanes 1 and 3 then you are seeing interaction.
your experimental design appears to be okay
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#3
PeterJallerup
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Posted 01 February 2012 - 03:55 AM
Thank you very much for the answer
I know a picture would have been awesome, but my supervisor have read 1984 too many times.
best regards
PJ
I know a picture would have been awesome, but my supervisor have read 1984 too many times.
best regards
PJ
#4
Adrian K
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Posted 01 February 2012 - 07:35 AM
PeterJallerup, on 01 February 2012 - 03:55 AM, said:
Thank you very much for the answer
I know a picture would have been awesome, but my supervisor have read 1984 too many times.
best regards
PJ
I know a picture would have been awesome, but my supervisor have read 1984 too many times.
best regards
PJ
Sorry, what you mean by "read 1984"? I'm not a native English speaker...
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
#5
mdfenko
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Posted 01 February 2012 - 09:37 AM
Adrian K, on 01 February 2012 - 07:35 AM, said:
Sorry, what you mean by "read 1984"? I'm not a native English speaker...
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
