BCA Instructions

BCA Reference

BCA:
1.    
Harvest sample into Rippa + Protease solution (big dishes – 100ul, combinations of wells from a plate – 50ul) (Protease breaks up the proteins into amino acides, measures total amount of amino acids)
2.    
Soncate for 10-15 sec each on 30% (except for GSIS
3.    
Centrifuge @ 12000xg, 4C, 10 min (this step doesn't apply to GSIS)
4.    
Dilute 3ul samples into 27ul ddh20
5.    
Mix Reagents 50:1 (A:B)
6.    
Rinse plate
7.    
190ul reagent mix +10ul samples (the order you add will be TBD by Amanda)
                - Prepare samples in duplicate
8.    
Add 10ul for you standards
9.    
Incubate for a minimum of 30 min (up to 2 hours
10.   Read in plate reader