Agrisera blog

During the recent 45th Annual Midwest/Southeast Photosynthesis Conference in Indiana, the Agrisera Best Talk Prize was awarded to Kelsey Dahlgren from the University of Colorado, Boulder. Kelsey works in Dr Jeffrey Cameron’s lab and gave an outstanding presentation entitled, "Development of a proximity-based proteomics technique using APEX2 to determine the thylakoid lumen proteome of Synechococcus sp. 7002." She is using both mass spectrometry and immunoblotting to characterize the lumenal proteome. 

Our warmest congratulations Kelsey. We are waiting your free antibody choice!

If no signal is detected in a total cell extract, it is worth fractionating, to obtain specific chloroplast, mitochondria or nuclei enriched fractions. This procedure will concentrate a target protein and lead to the detection of a desired band (for non-cytoplasmic proteins). This is shown in the example above - from no band in a total cell extract, to a specific band in photosystem I fraction.
This approach is especially important when working with proteins of low expression. Fractionation combined with using a specific organ, e.g. the root tip, or a leaf at a certain developmental stage, will greatly contribute to the successful identification of your protein of interest. 

Examples of protocols to fractionate can be found here.
For Agrisera Western Blot Guide and video tutorials, check here.
In case of any questions, you are always welcome to contact Agrisera Technical Support.