Lab 8 ELN Weekly Guidelines - Western Blot and Quantification
We are continuing the transition to study the new vector: pET41a-CaMPARI.
pET41a is a BACTERIAL EXPRESSION vector that also has a GST tag at the 3’ end of
CaMPARI. The theme of these labs is to isolate and visualize our CaMPARI protein (made
possible by the GST tag) and understand how CaMPARI functions as a calcium indicator.
Then we can finally move on to transfecting (aka ‘transforming’ but for mammalian cells)
our pHLsec-CaMPARI vector into mammalian cells and perform our calcium detection
experiment on different cancer drugs applied to our Chinese Hamster Ovary cells.
LADDER REFERENCE - Precision Plus Protein WesternC standards
After 5:50 pm (cutoff time for adding your individual data), copy it into your own separate
spreadsheet to perform average and SEM calculations. Don’t do your calculations in the
class spreadsheet!
Why are we analyzing class data?
The answer: Biological Replicates! We also want to practice calculating SEM
PURPOSE
We are continuing the transition to study the new vector: pET41a-CaMPARI.
pET41a is a BACTERIAL EXPRESSION vector that also has a GST tag at the 3’ end of
CaMPARI. The theme of these labs is to isolate and visualize our CaMPARI protein (made
possible by the GST tag) and understand how CaMPARI functions as a calcium indicator.
Then we can finally move on to transfecting (aka ‘transforming’ but for mammalian cells)
our pHLsec-CaMPARI vector into mammalian cells and perform our calcium detection
experiment on different cancer drugs applied to our Chinese Hamster Ovary cells.
LADDER REFERENCE - Precision Plus Protein WesternC standards
After 5:50 pm (cutoff time for adding your individual data), copy it into your own separate
spreadsheet to perform average and SEM calculations. Don’t do your calculations in the
class spreadsheet!
Why are we analyzing class data?
The answer: Biological Replicates! We also want to practice calculating SEM
PURPOSE
