Can i send protein band from SDS-PAGE for MAlDI......... - What could be the complication!!!!!!!!!! (Jan/01/2009 )
Hi all
Wishing you avery happy new year............
I want to share one prob with youl.................. and hope that i would definetly get some nice suggestion ............
I am Phd student, doing work on 2-DE. these days i want to finish off my work. As the facilitiess are limited.
i found conspicous difference in SDS profile as well as in 2 dimensional PAGE profile(among controland treated). But the prob is that as we send spot to another institute for MALDI-TOF. Their technician there are very fussy and do such a miss handling that eveytime complain for keratin. And my phd degree is waiting for on the MALDI data as the differece between 1-Dimensional is also very clear............
Can i send band from SDS PAGE profile for MALDI-TOF istead of 2 DE.......... to me i think i can............. if the difference is very much clear...................
has any body try it..............
kindly give me suggestion...........
waiting for your reply....
thanx in advance
Wishing you avery happy new year............
I want to share one prob with youl.................. and hope that i would definetly get some nice suggestion ............
I am Phd student, doing work on 2-DE. these days i want to finish off my work. As the facilitiess are limited.
i found conspicous difference in SDS profile as well as in 2 dimensional PAGE profile(among controland treated). But the prob is that as we send spot to another institute for MALDI-TOF. Their technician there are very fussy and do such a miss handling that eveytime complain for keratin. And my phd degree is waiting for on the MALDI data as the differece between 1-Dimensional is also very clear............
Can i send band from SDS PAGE profile for MALDI-TOF istead of 2 DE.......... to me i think i can............. if the difference is very much clear...................
has any body try it..............
kindly give me suggestion...........
waiting for your reply....
thanx in advance
do you mean a band from 1D gelelectrophoresis instead of 2D GE? It will be of high risk if it is below 100 kDa; you may get several signals which will not be a clear result/score if you match more than 1 or 2 polypeptides...
Hi
I do it regularly but only in cases where the protein conc. is very high and normally make a cut within the band (to avoid other bands). Also run a longer gel.
TC
Wishing you avery happy new year............
I want to share one prob with youl.................. and hope that i would definetly get some nice suggestion ............
I am Phd student, doing work on 2-DE. these days i want to finish off my work. As the facilitiess are limited.
i found conspicous difference in SDS profile as well as in 2 dimensional PAGE profile(among controland treated). But the prob is that as we send spot to another institute for MALDI-TOF. Their technician there are very fussy and do such a miss handling that eveytime complain for keratin. And my phd degree is waiting for on the MALDI data as the differece between 1-Dimensional is also very clear............
Can i send band from SDS PAGE profile for MALDI-TOF istead of 2 DE.......... to me i think i can............. if the difference is very much clear...................
has any body try it..............
kindly give me suggestion...........
waiting for your reply....
thanx in advance
You sure can, but if it is not long run and band is not fat enough, you will get a large number of proteins. No harm trying, unless it is like our institute where they charge $250/band. Insane..
Thanx so much for valuable suggestion................
What is the appropriate conc. of protein i should load on 1-DE so that i can send band for MALDI ? How is 35 microgram conc.
Wishing you avery happy new year............
I want to share one prob with youl.................. and hope that i would definetly get some nice suggestion ............
I am Phd student, doing work on 2-DE. these days i want to finish off my work. As the facilitiess are limited.
i found conspicous difference in SDS profile as well as in 2 dimensional PAGE profile(among controland treated). But the prob is that as we send spot to another institute for MALDI-TOF. Their technician there are very fussy and do such a miss handling that eveytime complain for keratin. And my phd degree is waiting for on the MALDI data as the differece between 1-Dimensional is also very clear............
Can i send band from SDS PAGE profile for MALDI-TOF istead of 2 DE.......... to me i think i can............. if the difference is very much clear...................
has any body try it..............
kindly give me suggestion...........
waiting for your reply....
thanx in advance
You sure can, but if it is not long run and band is not fat enough, you will get a large number of proteins. No harm trying, unless it is like our institute where they charge $250/band. Insane..
What is the appropriate conc. of protein i should load on 1-DE so that i can send band for MALDI ? How is 35 microgram conc.
Wishing you avery happy new year............
I want to share one prob with youl.................. and hope that i would definetly get some nice suggestion ............
I am Phd student, doing work on 2-DE. these days i want to finish off my work. As the facilitiess are limited.
i found conspicous difference in SDS profile as well as in 2 dimensional PAGE profile(among controland treated). But the prob is that as we send spot to another institute for MALDI-TOF. Their technician there are very fussy and do such a miss handling that eveytime complain for keratin. And my phd degree is waiting for on the MALDI data as the differece between 1-Dimensional is also very clear............
Can i send band from SDS PAGE profile for MALDI-TOF istead of 2 DE.......... to me i think i can............. if the difference is very much clear...................
has any body try it..............
kindly give me suggestion...........
waiting for your reply....
thanx in advance
You sure can, but if it is not long run and band is not fat enough, you will get a large number of proteins. No harm trying, unless it is like our institute where they charge $250/band. Insane..
I am supposed to send at leats ~50 pmoles