Western Blot Presentations

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