Molecular weight marker - got different pattern! (Nov/04/2009 )
Hi
i am using a molecular weight marker from Bangalore Genie (India)(Old catalog no. PMWM, new catalog no. 105979). In the image below A is the pattern which the company has given in the catalog, whereas B is the pattern which I got on same- 12% SDS-PAGE. As you can see clearly, there is a difference in the spacing between bands of the 2 patterns? Can anyone comment on why is it so? Is such case can I just count number of the bands and then compare the pattern order wise?
ram on Nov 4 2009, 05:49 PM said:
i am using a molecular weight marker from Bangalore Genie (India)(Old catalog no. PMWM, new catalog no. 105979). In the image below A is the pattern which the company has given in the catalog, whereas B is the pattern which I got on same- 12% SDS-PAGE. As you can see clearly, there is a difference in the spacing between bands of the 2 patterns? Can anyone comment on why is it so? Is such case can I just count number of the bands and then compare the pattern order wise?
Hi Ram,
The spacing depends on the gel percentage and for how long the gel was run....more the run time, more will be the resolution. Similarly on a lower percentage gel, the upper area will be expanded........to compare with ur gel, u will hv to count the no. of bands....
ram on Nov 4 2009, 07:19 AM said:
i am using a molecular weight marker from Bangalore Genie (India)(Old catalog no. PMWM, new catalog no. 105979). In the image below A is the pattern which the company has given in the catalog, whereas B is the pattern which I got on same- 12% SDS-PAGE. As you can see clearly, there is a difference in the spacing between bands of the 2 patterns? Can anyone comment on why is it so? Is such case can I just count number of the bands and then compare the pattern order wise?
Your spacing looks appropriate for a 12% SDS-PAGE gel, based on the molecular weights of the markers.
But both A and B are 12% PAG, then why is there such difference !?
Maybe the lot you received is different than what is printed in the catalog.
Is it just my imagination, or does band 3 on the right have a doublet?
hi ram... don mind but its bangalore genie!!!
this is not a surprice to me... and ya jsut possible that they boiled for less time you for more.. or something like that.. i mean sample treatent might hav been different!!!
or as lab rat says it might be a defective lot also!!!
i wuld not just count the bands and go ahead as the saing does not look okay to me given the molecular weights...
Yah.. there is a doublet in band # 3. If this is counted as 2 bands, then there are 6 bands in A and 7 in B!
Pradeep...does the inconstant pre-loading boiling really results in such a variable results?
ram on Nov 5 2009, 03:10 PM said:
Pradeep...does the inconstant pre-loading boiling really results in such a variable results?
well, i don't think so........such variability, due to boiling, i doubt???? y dont u try NEB ladder
DRN on Nov 5 2009, 03:12 PM said:
ram on Nov 5 2009, 03:10 PM said:
Pradeep...does the inconstant pre-loading boiling really results in such a variable results?
well, i don't think so........such variability, due to boiling, i doubt???? y dont u try NEB ladder
well i was telling that cause i have observed (not with marker) with some proteins, that under reduced conditions, even the boiling time lead to difference in the migration pattern!!!
But my stress was on the sample preparation.. boiking was jsut an example
DRN on Nov 5 2009, 12:42 AM said:
ram on Nov 5 2009, 03:10 PM said:
Pradeep...does the inconstant pre-loading boiling really results in such a variable results?
well, i don't think so........such variability, due to boiling, i doubt???? y dont u try NEB ladder
because I dont have it right now!