Archives

Fig. 1A. Crop the images to include only the area containing well-resolved protein spots. Use the Advanced Crop function to ensure that the cropped images are registered with each other. Place the crosshairs on a recognizable feature common to both images (Edit > Image > Advanced Crop > Define Crop Area).

Fig. 11. If desired, the match may be further refined by manually adding and deleting spots. Spot matches may be systematically verified using the Spot Review Tool (Analyze > Spot Review).

Fig. 10. Selecting Edit>Experiment Summary will display the corrected match rate.

Fig. 9. A more accurate match rate can be determined by deleting incorrectly identified spots. These can often be visually identified near the edges of the image. In this case, a series of spots along the top edge of the blot image probably do not correspond to real proteins. Spurious spots are deleted by selecting Edit > Spot > Remove Spot. The spots are erased by clicking on the spot or drawing a box around a set of spots. This operation should be performed both on the raw 2-D image and on the master.

Fig. 8. The resulting images will have identified spots marked with a colored “+” and matched spots marked with an “x” in a different color. This will aid in determining whether or not a spot identification or match is valid.

Fig. 7. To show matched spots, select Analyze>Analysis Set Manager,… then Create and Matching.

Fig. 6. Choosing View > Spot > Show Crosshairs or pressing F5 will show the detected spots.

Fig. 5. Match Rate 2 for the western blot image is the ratio of the number of protein spots recognized by the antibody to the total number of proteins detectable in the sample.