I run the marker and unknown protein, and i got the very clear band of the marker. The molecular weight of the marker are 10 - 250 kda. After making a graph for rf vs log MW, the equation is not linear, the r2 only 0,91. Could i use this equation to estimate the unknown protein? or , i have to use the other equation? Thanks for your kind attention and suggestion.
Can you share data for the same? I can give you xls sheet from which you can estimate..
Shaileh parab : i already attached the data in excell sheet... Here, i can see the data attached under the pic. I just take the linear graph part and make a new equation...
Your protein's mass is near the bottom end of the standards range. Try running it on a higher percentage acrylamide gel or a Tris-tricine gel and using standards with a lower range of molecular weights to get a better estimate.
Attached may help - use power function in Excel when adding trend line
www.malariaresearch.eu
I would run a 12.5% gel with a 14-97 kDa standard, or any other mid-range marker set.
I agree with Adam and Lillian. However I have attached file. you have have to insert rf of protein of interest in "Yellow"and following will be calculated with formulae.
thanks a lot to all comments. Thanks for helping to calculate the formula. Using the lower molecular weight standard will be give the better estimation, I agree with it :), hopefully i can find the standard. About the gel... i already run thhigher percentage of polyacrilamide, the result is the band stay thick , eventhough the other band which lower MW already separated. Tris tricine gel also will be helpfull for better estimation, but lack of tricine in my lab, trying find to other place or have to order it:).
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