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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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no bands in sds-page
Posted by: lalitha25 (IP Hidden, New member, 2)
Date: February 2, 2005 12:06PM
Hi,
I loaded some food extracts to see the proteins in the gel. I didn't get any bands. I can see some very lightly stained lines but they dont look like regular bands. I made sure that the extracts I used have a good concentration of proteins using Lawry-Folin assay I dont understand the reason. Is it because of low m.wt proteins or proteins are breaking down to small pieces which are not visible? please help ---lalitha
Re: no bands in sds-page
Posted by: lalitha25 (IP Hidden, New member, 2)
Date: February 3, 2005 08:49AM
actually when I used silver staining I can see smears instead of bands. do u have any ideas??
protein loaded
Posted by: huangz123 (IP Hidden, Junior member, 19)
Date: February 3, 2005 09:57AM
what is your food extract? It sounds to me like meat or maybe plant? Did you ever get nice bands with the same fresh extract with eough protein loaded? How much ug did you load your sample?
Re: protein loaded
Posted by: lalitha (IP Hidden, Unregistered user, )
Date: February 4, 2005 05:26AM
Vanilla is the extract - plant extract. For other plant extracts I get good bands . For this particular one i get smeaed zone between two vertical lines. I loaded 150 ug in 15% gel
Re: no bands in sds-page
Posted by: a.rhum (IP Hidden, Unregistered user, )
Date: February 8, 2005 05:06AM
I have done SDS PAGE for my protein samples and no bands has been shown on the gel is it because of the samples have been stored for long time nearly three weeks or because of the concentration
please helpe
Re: no bands in sds-page
Posted by: mamuller (IP Hidden, Junior member, 10)
Date: February 12, 2005 10:26PM
Hi,
This may be due to: 1) Staining solution bootle was open too long and evaporated 2) Bottle has been shelf stored tool long, staining solution need to be shaked for even mixe
Re: no bands in sds-page
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: March 15, 2005 03:42AM
Two suggestions:
1- Incubate your vanilla extract in Laemilli buffe for 5 minutes before boiling the sample. This will ensure bette protein extraction by the SDS in the loading buffer. 2- Vanilla extract might have interfring chemicals that gave you false protein concentrations using your Lawry-Folin assay, so I would suggest using BCA or Bradford assay. Good Luck!
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