I am interested in comparing the protein:lipid of pollen between three sister plant species.

This information will then be linked to pollinator communities and rate of visitation to each of the plant species.

In published literature it seems that the standard protocol is to use the Bradford protein assay for quantifying protein content and a modified protocol of lipid quantification from Van Handel et al. 1988. (Vaudo et al. 2020).

I will be following this standard method, but am curious as to why the BCA assay for protein quantification of pollen (Vanderplank et al. 2014) is not the standard?

From what I understand the methodology used in the BCA method quantifies the amount of polypeptides which are the most nutritionally relevant units of protein in relation to insects.

When breaking the pollen cell walls the Bradford protein assay uses a NaOH. Which types of protein I would be capturing with this sort of methodology?

I am guessing that the Bradford assay is the standard because and the goal is to calculate a ratio, not to report the exact quantities or identify the types of proteins. Also, it appears to be an easier methodology.

Thank you!

Van Handel, E., & Day, J. F. (1988). Assay of lipids glycogen and sugars in individual mosquitoes: correlations with wing length in field collected Aedes vexans. Journal of American Mosquito Control Association, 4(4), 549–550.

Vanderplanck, M. Leroy, B. , Wathelet, B. W., & Wattiez, R. W. (2014). Standardized protocol to evaluate pollen polypeptides as bee food source Original article Standardized protocol to evaluate pollen polypeptides as bee food source, (March). https://doi.org/10.1007/s13592-013-0239-0

Vaudo, A. D., Tooker, J. F., Patch, H. M., Biddinger, D. J., Coccia, M., Crone, M. K., … Grozinger, C. M. (2020). Pollen Protein : Lipid Macronutrient Ratios May Guide Broad Patterns of Bee Species Floral Preferences. https://doi.org/10.3390/insects11020132