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There have been 4 items by s056870 (Search limited from 22-September 19)


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#111042 Converging lanes in Western

Posted by s056870 on 26 May 2011 - 01:30 PM in SDS-PAGE and Western Blotting

Hi mdfenko,

Thank you for the reply.

Group 2 was treated with a flavonoid-rich food extract, while group 1 was a control in which the cells were incubated in cell culture medium only for the same period of time.

Regards,
Stephanie


The proteins from two different treatment groups were extracted using exactly the same protocol and reagents. Do you reckon it might be the treatment that made the difference? However, the problem occurred in both groups, although the majority is in one of them.

it depends on the treatment. what was the treatment?

also, were both groups treated equally or was there a difference in treatment?




#110585 Converging lanes in Western

Posted by s056870 on 23 May 2011 - 02:14 AM in SDS-PAGE and Western Blotting

Hi mdfenko,

Thanks for your reply.

The proteins from two different treatment groups were extracted using exactly the same protocol and reagents. Do you reckon it might be the treatment that made the difference? However, the problem occurred in both groups, although the majority is in one of them.

P.S. The lanes on the films are (from the left) group 1, 2, 1, 2, 1, 2, 1, 2.

Thanks,
Stephanie

were they all extracted using the same buffer? detergents?

the effect you are seeing can be caused (not exclusively) by detergents or other additives to the buffer or lipids (from the sample).




#110166 Converging lanes in Western

Posted by s056870 on 18 May 2011 - 06:48 AM in SDS-PAGE and Western Blotting

Hi Buena,

Thanks for your reply.

As I understand, you seem to regard the wider bands as abnormal rather than the narrower ones, is that correct? But the thing is the wide bands have the same width as the wells for protein loading, while the narrow bands are resulted from converging lanes when the gel was being run. So, I'm not sure if I agree with your ideas.

Regarding ionic strength of the samples, I'm afraid I've never checked that. But all the protein samples were extracted from Caco-2 cells.

Thanks again for your comments.

Regards,
Stephanie



Dear all,

I wonder if anybody has seen this phenomenon.
Attached File  images for bioforum.bmp   831.85KB   784 downloads
Some of the lanes in the gels converged to become very narrow when I ran the SDS-PAGE. The bands detected were then very small. It also seemed that the bands in the narrow lanes shifted a bit upwards than the more normal lanes. Could somebody please suggest any reasons for this weird result?

Steph

Hola, for me the wide bands are due to an salt excess, as the used buffer for wide bands has more ionic strength than the narrow´s. An example, if you analized the elution profile of an ion exchanger the bands width is increasing as the salt concentration arises. Have the your samples different ionic strength?
Buena suerte




#110158 Converging lanes in Western

Posted by s056870 on 18 May 2011 - 03:06 AM in SDS-PAGE and Western Blotting

Dear all,

I wonder if anybody has seen this phenomenon.
Attached File  images for bioforum.bmp   831.85KB   784 downloads
Some of the lanes in the gels converged to become very narrow when I ran the SDS-PAGE. The bands detected were then very small. It also seemed that the bands in the narrow lanes shifted a bit upwards than the more normal lanes. Could somebody please suggest any reasons for this weird result?

Steph




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