Development of Western Blots for Actin without the use of radioactivity. Geoff Theobald STEP Summer Internship Program June 2003. Objective. To create a Western Blot without the use of radioactivity. Significance. Safety & disposal
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STEP Summer Internship Program
To create a Western Blot without the use of radioactivity.
Pic. Of membrane radioactivityWestern Blot
Actin ProteinProbing with an antibody to actin
Probe blot with antibody
Secondary Antibody for Rabbit
= Primary Antibody
= Secondary Antibody
Primary Rabbit Antibody for Actin
= DDAO phosphate
= Alkaline phosphatase
Western BlotDetection by fluorescence
Pic. of Blot exposed to UV light radioactivityResults: detection with UV light
Pic. Of blot exposed to white light radioactivityResults: detection with white light
Pic of dot blot with all actin antibody radioactivity
Pic of membrane with all actin antibodyChemiluminescence assay
I would like to thank each and everyone of you for all of your advice, support, and fun I had these past two weeks.