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SDS PAGE Reagents

This document provides instructions for making stock solutions and concentrated buffers for SDS-PAGE gel electrophoresis, including a 20% SDS stock solution, 1.5M and 0.5M Tris stock solutions at pH 8.8 and 6.8, respectively, 4X resolving and stacking buffers, 10X running buffer, and a 2X SDS-PAGE sample buffer.

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338 views1 page

SDS PAGE Reagents

This document provides instructions for making stock solutions and concentrated buffers for SDS-PAGE gel electrophoresis, including a 20% SDS stock solution, 1.5M and 0.5M Tris stock solutions at pH 8.8 and 6.8, respectively, 4X resolving and stacking buffers, 10X running buffer, and a 2X SDS-PAGE sample buffer.

Uploaded by

Peer Latif
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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SDS-PAGE

Reagents

Stock solutions:

20% SDS
20 g SDS in 100 ml final volume of dd H2O

1.5 M Tris, pH 8.8


27.23 g Tris base + 80 ml dd H2O
Adjust to pH 8.8 with 6N HCl, bring to 150 ml final volume with ddH2O.

0.5 M Tris, pH 6.8


6 g Tris base + 60 ml dd H2O
Adjust to pH 6.8, bring to 100 ml final volume with ddH2O.

Concentrated buffers:

4X resolving buffer
1.5 M Tris base, 0.4% SDS, pH 8.8
Mix 10 ml 1.5 M Tris, pH 8.8 + 0.2 ml 20% SDS

4X stacking buffer
0.5 M Tris, pH 6.8, 0.4% SDS
Mix 5 ml 0.5 M Tris, pH 6.8 + 0.1 ml 20% SDS

10X running (electrode or reservoir) buffer


0.25M Tris, 1.92 M glycine, 1% SDS, pH 8.3.
Mix 30 g Tris base and 144 g glycine per liter

2X SDS-PAGE sample buffer


Mix the following reagents:
0.5 M Tris, pH 6.8 3.50 ml
Glycerol, 50% 4.00 ml
β-mercaptoethanol 0.4 ml (in hood!)
20% SDS 0.2 ml
bromophenol blue 50 mg
Bring to 10 ml volume with ddH2O.

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