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Scanning and Analyzing Western Blots Using LiCor Odyssey

5 bytes added, 15:51, 26 March 2021
→‎Analysis/Quantification
#Click '''Redraw Boundaries''' and drab the boundary to include all lanes (not ladder(s)).
#To select the bands of interest, select the '''700 or 800 channel''' by clicking the '''X''' box next to the channel on the right side of the screen (with the vertical '''Display''' tab selected). Add boxes to your bands (see below) and repeat for each channel. Each band of interest should have one box around it.
*##You can use '''Add''' in the '''Bands''' menu in the top center menu bar to add boxes around individual bands to be quantified. *##Or you can use '''Add to All Lanes''' in the '''Bands''' menu to add bands of one size across all lanes. This is much faster if you have many lanes.
#Click '''Select''' in the '''Bands''' menu.
#Click the vertical '''Profiles''' tab (below the '''Display''' tab. Adjust the edges of the boxes around each band to include the entire band using the graphs. Each edge should be on the axis and should capture the desired peak(s).
#Once all bands are adjusted, click the '''Western Bands''' tab within the table at the bottom of the screen.
#Click the triangle in the top left corner of the table to copy the table (or the lanes you want). Copy into an Excel spreadsheet or a Text file to use in Excel on your own computer.
*##Use this format: yy_mm_dd_nameofexperiment_quantification
#Save your work/'''Work Area''' within '''ImageStudio''' and save pictures (without features) to your flash drive. You can remove the features by unchecking the boxes in the '''Show''' menu bar on the right of the top menu bar.
*##For the former, click the '''disk icon''' at the top the screen.*##For the latter, click the yellow '''IS''' circle at the top left of the screen and select '''Export''' and '''Image for Print'''. Save as '''TIFF''', '''PNG''', or '''PDF''' as needed. (Note: Save pictures in greyscale by clicking the greyscale box in the '''Display''' tab above '''Profiles''' (see #6).*##You can also save everything in to a '''Lab Notebook''' as a PDF using the '''Lab Book''' tab in the top menu.
Note: To quantify the total protein scan, use steps 4-7, but with adjustments to 7.
#Control+Click to select all bands in one lane and click '''Merge''' in the '''Bands''' menu to create one box around the lane.
#Adjust the edges, as above.
#Use the '''Western Bands'''' tab, as above to copy the quantification values (see #109).
==Finishing Up==
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