Measuring protein concentration and sample denaturation

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  • čas přidán 28. 08. 2024
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    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.

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