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00:19
1.
Western Blot - 2_Electrophoresis [Video from GeneTex]
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00:12
2.
Clean the glass plates
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00:22
3.
Assemble the gel apparatus
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00:08
4.
Leaking test
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00:08
4.1
Pour off H2O
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00:12
5.
Preparation of separating gel
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00:08
5.1
Add H2O
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00:10
5.2
Add 1.5M Tris (pH8.8)
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00:11
5.3
Add 30% polyacrylamide
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00:13
5.4
Add 10% SDS
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00:13
5.5
Add 10% ammonium persulfate (APS)
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00:17
5.6
Add TEMED
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00:11
5.7
Add the mixture into the gap between the glass plates
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00:05
6.
Overlay with 75% EtOH
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00:00
6.1
Remove 75% EtOH
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00:03
6.2
Remove 75% EtOH
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00:04
7.
Waiting for 30-60 min
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00:00
8.
Preparation of stacking gel
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00:20
8.1
Add H2O
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00:07
8.2
Add 1M Tris (pH6.8)
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00:11
8.3
Add 30% polyacrylamide
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00:10
8.4
Add 10% SDS
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00:11
8.5
Add 10% ammonium persulfate (APS)
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00:15
8.6
Add TEMED
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00:08
8.7
Add the mixture into the gap between the glass plates
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00:16
9.
Insert a clean comb into the stacking gel solution
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00:04
10.
Waiting for 15-30 min
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00:17
11.
Remove the comb gently
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00:00
12.
Running the gel
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00:20
12.1
Assemble the electrophoresis apparatus
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00:07
12.2
Add running buffer
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00:10
12.3
Vortex samples & protein marker
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00:03
12.4
Centrifuge samples & protein marker
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00:15
12.5
Loading samples & protein marker
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00:05
12.6
Attach the electrophoresis apparatus to the power supply
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00:06
12.7
Apply the voltage of 80V to the gel, run 15 min
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00:16
12.8
After the dye front has moved into the stacking gel, increase the voltage to 140V and run 50 min