Hello dears.
I did run SDS-PAGE gel yesterday, after staining the gel and destaining the gel i saw that protein's bands did not migrate to the
end of the gel and they have just migrated half of the gel and protein marker too.i don't know why it is happening to my gels.
I have changed my buffers and made a fresh buffer and acrylamide too and then run the gel but the same result obtained to my gel.
Could you please help me with this? what is the reason of this problem? it has not been occurred to me since i have started my job.my protein molecular weight is 38-39kD and i don't know if i have my protein presented on this gel or not.
Do i have my protein band on lane 1 and 3?
lane 1 and 3 are T4 (4 hours after induction) and lane 2 and 4 are T0 (before induction).
The protein marker that i've used is Thermoscientific Unstained Protein Molecular Weight Marker 26610.
Protocol i have used for SDS-PAGE buffers and acrylamide obtained from "Short Protocol in Cell Biology" book,is it a good
source for protocol?
i have attached protocol and gel pictures.
Thanks in advance.
Hi there,
Have you tried running the gel for a longer period of time to increase the distance the proteins migrate?
Thanks dear Rory Little.
No i have not try it yet.
when ever the stained protein samples come out of the bottom of the gel i turn off the power system(Biorad Power Supply system). i use voltage of 85 v and my gel concentration is 15%.
Should i let the samples migrate after the stain comes out?
Yeah, it sounds like you need to keep running the gel after the dye comes out the bottom. You could also try increasing the voltage you run the gel at (100-200V). If you can get some this protein ladder is really good: http://www.bio-rad.com/en-ch/sku/1610374-precision-plus-protein-dual-color-standards-500-mul, as you can see the ladder migrate while the gel is running.
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