I recently completed a experiment on SDS-Page electrophoresis to identify the molecular weight of my unknown proteins (unknown a and unknown b). I recently received my photograph that I took and I am having a hard time reading it. First off I am having difficult interpreting my photo as I only know that the 5th line is the marker and on top of that I do not know how to make a plot of the log of known protein MW versus gel mobility from my data and used it to estimate the molecular weight of my unknown proteins. The MW of known proteins are phosphorylase:97400Da bovine serum albumin:66200Da ovalbumin 45000Da carbonic anhydrase:31000Da soybean trypsin inhibitor:21500Da and lysozyme:14400Da all at 1mg/ml.
If you could help me out on these two aspects (how to read the photo and making the graph) I would greatly apprecited it.
Attach is my photo and my biorad marker index and also the lab instructions. Also if more information is needed please let me know and I will give it to you.
The lane markers are loaded as follows:
Lanes 1,2,3= Unknown (A)- a protein that didn't resolve on my gel
Lanes 4,5,6= Unknown (B)- is the other protein that that I loaded that did resolve
Lane 8= marker
Please ignore lanes 7,9,10
First of all, your MW standards that you list above is irrelevant as it was not the set of standards you actually loaded in lane 8. Rather, lane 8 contains the Biorad Marker Index standards that you have shown in the picture below the photograph of the gel.
That's probably why you were having trouble. If you look at the Biorad Marker Index picture and compare it to lane 8 you will quickly see that the two are one and the same. In fact, you can even see the yellow protein marker at 10 kD at the bottom of the gel.
First, print the document containing the ...
The expert examines SDA-Page electrophoreis to identify the molecular weigh.