Bitan:densitometry: Difference between revisions
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(New page: The signal from a Western Blot can be quantified using ImageJ. Here is how to do lane-by-lane densitometry of a Western Blot.<br> =ImageJ= ImageJ is an image processing software package m...) |
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The signal from a Western Blot can be quantified using ImageJ. Here is how to do lane-by-lane densitometry of a | The signal from a Western Blot (WB) can be quantified using ImageJ. Here is how to do lane-by-lane densitometry of a WB.<br> | ||
=ImageJ= | =Setup ImageJ= | ||
ImageJ is an image processing software package maintained by the NIH, you can get it [http://rsbweb.nih.gov/ij/download.html here].<br> | ImageJ is an image processing software package maintained by the NIH, you can get it [http://rsbweb.nih.gov/ij/download.html here].<br> | ||
If you want to estimate the molecular weights of any of the bands in your blot you will need two things:<br> | If you want to estimate the molecular weights of any of the bands in your blot you will need two things:<br> | ||
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#copy the molwt.txt file into the macro folder of ImageJ (/Applications/ImageJ/Macros)<br> | #copy the molwt.txt file into the macro folder of ImageJ (/Applications/ImageJ/Macros)<br> | ||
#Run ImageJ<br> | #Run ImageJ<br> | ||
#In ImageJ | #In ImageJ do Plugins->Macros->Install<br> | ||
#Select molwt.txt from the ImageJ/Macros folder<br> | #Select molwt.txt from the ImageJ/Macros folder<br> | ||
=Using ImageJ to Plot WB Signal= | |||
#Launch ImageJ and open an image of a WB.<br> | |||
#The blot's lanes may not be oriented vertically, do Image->Rotate->Arbitrarily... in 0.5 or 1.0 degree increments.<br> | |||
#Use the palate to select the "Rectangular selections" tool.<br> | |||
#Select an entire lane at once with the Rectangular selections tool.<br> | |||
#Do Analyze->Gels->Select First Lane (Cmd+1).<br> | |||
#Use the left or right arrow key to move the highlighted box to select the next lane.<br> | |||
#Do Analyze->Gels->Select Next Lane (Cmd+2).<br> | |||
#Repeat the previous two steps until all lanes have been selected.<br> | |||
#Do Analyze->Gels->Plot Lanes.<br> | |||
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Revision as of 10:32, 7 December 2009
The signal from a Western Blot (WB) can be quantified using ImageJ. Here is how to do lane-by-lane densitometry of a WB.
Setup ImageJ
ImageJ is an image processing software package maintained by the NIH, you can get it here.
If you want to estimate the molecular weights of any of the bands in your blot you will need two things:
- a text file of the molecular weights of the ladder
- this molecular weight macro
Install the molecular weight macro:
- copy the molwt.txt file into the macro folder of ImageJ (/Applications/ImageJ/Macros)
- Run ImageJ
- In ImageJ do Plugins->Macros->Install
- Select molwt.txt from the ImageJ/Macros folder
Using ImageJ to Plot WB Signal
- Launch ImageJ and open an image of a WB.
- The blot's lanes may not be oriented vertically, do Image->Rotate->Arbitrarily... in 0.5 or 1.0 degree increments.
- Use the palate to select the "Rectangular selections" tool.
- Select an entire lane at once with the Rectangular selections tool.
- Do Analyze->Gels->Select First Lane (Cmd+1).
- Use the left or right arrow key to move the highlighted box to select the next lane.
- Do Analyze->Gels->Select Next Lane (Cmd+2).
- Repeat the previous two steps until all lanes have been selected.
- Do Analyze->Gels->Plot Lanes.
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