Agrisera blog

Agrisera on 45th Annual Midwest/Southeast Photosynthesis Conference

45th Annual Midwest Southeas Photosynthetic Conference

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!

Read more 2019-10-30

Common misconceptions about antibodies

Can the same protocol can be applied to all antibodies?

No, this is NOT the case.

Each protein-antibody pair may require slightly modified binding and detection conditions. The variations may for example be a membrane with another pore size and physical properties (nitrocellulose or PVDF) as well as different blockers, incubation times and primary antibody dilutions. Some antibodies may work better in TBS-T buffer, others with PBS-T.

These issues are disussed in Agrisera Western blot guide, which we recommend you to keep in your lab!

Western Blot Guide
Get your free copy of the Agrisera Western Blot Guide here!

Read more 2019-10-21

Why fractionate?

Advantages of fractionation

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

Read more 2019-10-14

Is antibody always to blame?

Western blot using anti-BAK1 antibodies

In the example above, the same antibody was used (Anti-BAK1, AS12 1858) but the proteins were extracted with two different buffers.
Minor adjustments in the buffer components resulted in the improved detection of the target protein - from NO band to a band of correct MW. 

If you are interested in more tricks to optimize your western blot, you are welcome to contact us or request a copy of the Agrisera Western Blot Guide.
Read more 2019-10-07
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