If you have questions or comments please go to the new channel Ciencia y Science and ask your questions there so that I can answer them. https://www.youtube.com/channel/UCvOlUnIRNhI2PonooNtcJ7w
In this tutorial video I have included both 1) measuring protein concentration from sample lysates using the Pierce BCA protein assay and 2) Denaturing sample lysates using 4X Laemmli buffer
Time stamp
0:00 Introduction
0:16 Pierce BCA protein assay from Thermo
0:43 Protocol/instructions from kit
1:42 Albumin standard I use to make my protein standards from Thermo
2:18 Loading protein standards and lysate samples into 96-well plate
11:11 Calculations for BCA reagents A and B
14:10 Loading 200uL of reagents A and B mixture
16:10 Incubate plate at 37C for 30 minutes
16:29 Read plate at 562nm using Cytation 5
19:46 Saving your data in excel
22:09 Calculations to determine sample volume for denaturation
27:01 Bio-Rad 4X Laemmli Sample Buffer instructions
29:40 Preparing sample lysates with 4X Laemmli buffer
33:48 Vortex samples, quick spin and heat denaturation at 90C.
