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Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


how to measure and identify my protein after western blotting, help me

how to measure and identify my protein after western blotting, help me - Western Blot Forum

how to measure and identify my protein after western blotting, help me - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 02-22-2010, 12:27 AM
Pipette Filler
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Exclamation how to measure and identify my protein after western blotting, help me



hi
i am working with PGT3 .am trying to increase the expression of protein using ecoli,and my treatments are two. well growing ecoli in betain and sorbitol and the other treatment without betain and sorbitol.
can u help me?i am done with my western blotting but cant figure out how to identify the sizes and bands i am seeing. i have been told to draw a graph,i am not understanding the whole process,pls help me,i am new in this are of science,nobody seems to nderstand this, i am sick of grad sch i wanna quit
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Old 06-02-2010, 05:49 PM
Pipette Filler
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Default Re: how to measure and identify my protein after western blotting, help me

Hi,

Don't panic.Be cooooooool.
So u are extracting protein from whole cell lyastes and loading them to SDS-PAGE gel and blotting onto the membrane and trying to detect ptn of interest.
Now u have to identify your ptn band......
First of all, are u using or running a ptn marker simultaneously along with ur sample?
If so then just compare the scale or length of the marker to ur sample and identify the band of ur interest(u should know the molecular weight of ur ptn)
If u want to quantify the ptn amount then reprobe the membrane with and internal control,measure the intensities of bands using image analysis software and try to get the ptn amount.......I hope this would be helpful

Quitting doesn't help ..............
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