When presenting a western blot in a Starr lab meeting or presentation, include the following information:
Title: Date, protein(s) and cell lysates including conditions being analyzed.
Subtitle: Your initials, and date where details can be found in your lab book (see Lab Book Details).
Original blot images
- Thumbnails of the entire images of the blots
- Filenames of the original images on proteinsimple machine
Blot image used for densitometry measurements
- Label every lane (optional: use numbers and have a number key below)
- Label ladder band sizes
- Label band(s) of interest with protein name
- Label control band with protein name
Densitometry Graph
- Title: Descriptive of experiment
- Label X-axis: Cells
- Label Y-axis: Protein expression relative to control lysate normalized to housekeeping gene. (change words in red words to your specific experiment)
Caption and conclusions
- Figure caption: Short paragraph describing Western Blot method.
- Figure conclusion: Short paragraph with your conclusions.
Transferring images to powerpoint using FIJI (ImageJ)
- Open the blot image and the ladder image and convert both images to 8 bit (Image > Type > 8-bit)
- Generally invert the blot image so that bands are black and background is white (Edit > Invert)
- Adjust both the blot image and the ladder image (Image > Adjust > Brightness/Contrast) so that they look good to you
- Rotate the images so the bands are in a horizontal line (Image > Transform > Rotate) then adjust Angle until bands look horizontal. Use Grid Lines = 20, Bilinear interpolation and click the preview box. Be sure to rotate both images exactly the same
- Clear everything outside of the ladder in the ladder image by drawing a rectangle around the ladder lane then use Edit > Clear Outside
- Using Screen Capture (shift-ctrl-command-4), take a snapshot of both the ladder image and the blot image and paste them into powerpoint.
- To position the ladder, return to FIJI and click on the blot image, then overlay the ladder (Image > Overlay > Add Image). Select the ladder image and set the opacity to ~75%. This will produce an image that will allow you to align position your ladder image next to your blot image in powerpoint.
Below is an example of a well-presented Western blot
