Date
2-13-2020
Faculty Mentor
Roger Sauterer, Biology
Loading...
Files
Download (59.0 MB)
Submission Type
Poster
Location
Houston Cole Library, 11th Floor | 4:00-4:10 p.m.
Description
Mammalian histones are highly alkaline proteins found in the nuclei that organize DNA into chromosomes and regulate transcription. We are investigating the interactions between histones and mitochondria by using cell fractionation and Western Blotting to identify the histones. The blocking step in the process coats the membrane with proteins or other molecules to reduce non-specific binding of the antibodies. We use 5% nonfat dry milk as a control for all experiments, it is essential to use this variation of milk because it reduces background noise and helps produce good, clear bands. Although 5% nonfat dry milk is widely used as a blocking buffer, we are testing different blocking buffers to see if the signal strength increases or completely strips proteins off of the blot, the conclusion of which blocking buffer works the best will be determined by the protein being examined. Since antibodies to histone H2A and H2B are low-affinity and give a weak signal, we are therefore testing different blocking buffers, such as milk concentrations ranging from 1 to 5%, BSA ranging from 1 to 5%, and hemoglobin ranging from 1 to 5%, as well as gelatin at 3%, and PVP or PEG individually ranging from 1 to 4% and different combinations of the aforementioned blocking buffers to determine which blocking buffer gives the best detection signal. Our preliminary results indicated that RVR/REG combinations provided the strongest signal, while gelatin tends to strip the proteins completely off of the blot and reduce the signal.
Keywords
student presentations, student posters, histones, blocking buffers
Rights
This video is the property of Jacksonville State University and is intended for non-commercial use. Video and images may be copied for personal use, research, teaching or any "fair use" as defined by copyright law. Users are asked to acknowledge Jacksonville State University. For more information, please contact digitalcommons@jsu.edu.
Disciplines
Biology
Recommended Citation
Tavis, Peighton and Harris, Shelby, "Blocking Buffers and Their Effects on Mammalian Histones" (2020). JSU Student Symposium 2020. 30.
https://digitalcommons.jsu.edu/ce_jsustudentsymp_2020/30
Presentation Information
Tavis, P. and Harris, S. (2020, 13 February). Blocking Buffers and their effects on mammalian histones. Poster presented at the 2020 JSU Student Symposium, Jacksonville State University, Jacksonville, AL.