Help! SDS-PAGE interpretation - (Mar/03/2007 )
Hi everyone,
Please excuse my ignorance, I'm a college student and have never worked with this stuff before. Thanks in advance for any and all advice.
I ran an SDS-PAGE gel and then stained with Coomassie blue. In the attached picture of the gel, (from left to right) the first two lanes are bacteria + IPTG + chloramphenicol, the next three are bacteria + IPTG, and the last three are control (uninduced bacteria).
Anyway, I'm having trouble interpreting the gel. Its my understanding that I need to measure from the top of the gel to the most prominent band in the lane and then use this data construct the graph. However it appears to me that though some lanes have more intense bands than others, the most prominent band appears at the same place in each lane (that is at 100 Mr corresponding to the marker just above the 70 Mr orange standard). That wouldn't make sense though...the graph would be meaningless. 
Please help.
You were told to construct a graph? What kind of graph?
what kind of graph do you want to make?
B)-->
Please excuse my ignorance, I'm a college student and have never worked with this stuff before. Thanks in advance for any and all advice.
I ran an SDS-PAGE gel and then stained with Coomassie blue. In the attached picture of the gel, (from left to right) the first two lanes are bacteria + IPTG + chloramphenicol, the next three are bacteria + IPTG, and the last three are control (uninduced bacteria).
Anyway, I'm having trouble interpreting the gel. Its my understanding that I need to measure from the top of the gel to the most prominent band in the lane and then use this data construct the graph. However it appears to me that though some lanes have more intense bands than others, the most prominent band appears at the same place in each lane (that is at 100 Mr corresponding to the marker just above the 70 Mr orange standard). That wouldn't make sense though...the graph would be meaningless.
Please help.[/quote] B)-->
Please excuse my ignorance, I'm a college student and have never worked with this stuff before. Thanks in advance for any and all advice.
I ran an SDS-PAGE gel and then stained with Coomassie blue. In the attached picture of the gel, (from left to right) the first two lanes are bacteria + IPTG + chloramphenicol, the next three are bacteria + IPTG, and the last three are control (uninduced bacteria).
Anyway, I'm having trouble interpreting the gel. Its my understanding that I need to measure from the top of the gel to the most prominent band in the lane and then use this data construct the graph. However it appears to me that though some lanes have more intense bands than others, the most prominent band appears at the same place in each lane (that is at 100 Mr corresponding to the marker just above the 70 Mr orange standard). That wouldn't make sense though...the graph would be meaningless.
Please help.[/quote]do you mean a Neville plot which refer to plotting Mw against Rf; intensity however is unaccounted...
Is it called the Neville plot? But I think you are right about plotting Mw against Rf..... just the migrating rate But this graph only determine the size of the protein isnt it?
