Title: Beefing up an Enzyme Lab: Acetylcholinesterase Activity in Extracts from Bean Beetles Standardized for Protein Content
Url: http://www.ableweb.org/volumes/vol-37/v37reprint.php?ch=73
Author: Hector Fermin
Fardad Firooznia
Avaje Jackson
Author's Address: City College of New York, Department of Biology, MR-526,160 Convent Ave. New York, NY 10031 USA
Email Address: hector@sci.ccny.cuny.edu
Description: The insecticide malaoxon inhibits acetylcholinesterase (AChE) activity. Various factors might
affect the effectiveness of malaoxon. We have previously described a colorimetric enzyme assay
to look at the differences in inhibition of AChE caused by malaoxon in crude extracts from the
bean beetle Callosobruchus maculatus in a set up that lends itself to a multi-week lab sequence.
Here we expand this set of exercises to include the Bradford assay to measure protein
concentration in crude extracts using a standard curve. This allows us to standardize the initial
activity of AChE based on protein content and thus consider both initial activity of the enzyme
and relative inhibition of the activity due to the insecticide. These two parameters have been
shown to be inversely correlated and may respond differently to environmental factors such as
the food source on which the animals are raised. These types of projects not only introduce
students to different biochemical techniques, but also raise questions such as costs and benefits
of insecticide resistance.
Keywords: bean beetle
enzyme assay
Topic: Animal Physiology
BEN Subject/Discipline Taxonomy: Biochemistry
Learning Resource Type: Laboratory exercise (DCMI Type Vocabulary)
Context: Undergraduate lower division 13-14
Format: pdf
Access Rights: This resource is for ABLE members only.
Rights: http://www.ableweb.org/copyright-policy/
Conference Location & Year: Boston University (2015)
Cumulative Rating: NOT YET RATED
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