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SDS-PAGE gel recipes
Author: Graeme J Gowans, updated date: , view: 32, Q&A: 0
Tags: Protein and SDS-PAGE

Procedure

SDS-PAGE: enough for 2 x mini gels (Biorad system):


10 ml of 15% separating gel            4 ml of 6% stacking gel

2.4 ml H2O                                      2.7 ml H2O

5 ml acrylamide (30 %)                  0.8 ml acrylamide (30 %)

2.5 ml 1.5 M tris (pH 8.8)               0.5 ml 1 M tris (pH 6.8)

50 µl SDS (20%)                            20 µl SDS (20%)

100 µl APS (10%)                          40 µl APS (10%)

10 µl TEMED                                 4 µl TEMED


10 ml of 13.5% separating gel        4 ml of 6% stacking gel

2.9 ml H2O                                    2.7 ml H2O

4.5 ml acrylamide (30 %)             0.8 ml acrylamide (30 %)

2.5 ml 1.5 M tris (pH 8.8)             0.5 ml 1 M tris (pH 6.8)

50 µl SDS (20%)                         20 µl SDS (20%)

100 µl APS (10%)                       40 µl APS (10%)

10 µl TEMED                              4 µl TEMED


10 ml of 12% separating gel            4 ml of 6% stacking gel

3.4 ml H2O                                2.7 ml H2O

4 ml acrylamide (30 %)            0.8 ml acrylamide (30 %)

2.5 ml 1.5 M tris (pH 8.8)         0.5 ml 1 M tris (pH 6.8)

50 µl SDS (20%)                      20 µl SDS (20%)

100 µl APS (10%)                   40 µl APS (10%)

10 µl TEMED                           4 µl TEMED


10 ml of 10% separating gel            4 ml of 6% stacking gel

4.1 ml H2O                                   2.7 ml H2O

3.3 ml acrylamide (30 %)            0.8 ml acrylamide (30 %)

2.5 ml 1.5 M tris (pH 8.8)            0.5 ml 1 M tris (pH 6.8)

50 µl SDS (20%)                        20 µl SDS (20%)

100 µl APS (10%)                      40 µl APS (10%)

10 µl TEMED                             4 µl TEMED


10 ml of 8% separating gel            4 ml of 6% stacking gel

4.7 ml H2O                                    2.7 ml H2O

2.7 ml acrylamide (30 %)             0.8 ml acrylamide (30 %)

2.5 ml 1.5 M tris (pH 8.8)             0.5 ml 1 M tris (pH 6.8)

50 µl SDS (20%)                         20 µl SDS (20%)

100 µl APS (10%)                      40 µl APS (10%)

10 µl TEMED                              4 µl TEMED





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