• 00:00 1.
    Western Blot - 2_Electrophoresis [Video from GeneTex]
  • 00:19 2.
    Clean the glass plates
  • 00:32 3.
    Assemble the gel apparatus
  • 00:54 4.
    Leaking test
  • 01:03 4.1
    Pour off H2O
  • 01:11 5.
    Preparation of separating gel
  • 01:23 5.1
    Add H2O
  • 01:32 5.2
    Add 1.5M Tris (pH8.8)
  • 01:43 5.3
    Add 30% polyacrylamide
  • 01:54 5.4
    Add 10% SDS
  • 02:08 5.5
    Add 10% ammonium persulfate (APS)
  • 02:22 5.6
    Add TEMED
  • 02:40 5.7
    Add the mixture into the gap between the glass plates
  • 02:51 6.
    Overlay with 75% EtOH
  • 02:57 6.1
    Remove 75% EtOH
  • 02:57 6.2
    Remove 75% EtOH
  • 03:01 7.
    Waiting for 30-60 min
  • 03:05 8.
    Preparation of stacking gel
  • 03:05 8.1
    Add H2O
  • 03:25 8.2
    Add 1M Tris (pH6.8)
  • 03:33 8.3
    Add 30% polyacrylamide
  • 03:45 8.4
    Add 10% SDS
  • 03:55 8.5
    Add 10% ammonium persulfate (APS)
  • 04:07 8.6
    Add TEMED
  • 04:23 8.7
    Add the mixture into the gap between the glass plates
  • 04:31 9.
    Insert a clean comb into the stacking gel solution
  • 04:47 10.
    Waiting for 15-30 min
  • 04:52 11.
    Remove the comb gently
  • 05:09 12.
    Running the gel
  • 05:09 12.1
    Assemble the electrophoresis apparatus
  • 05:29 12.2
    Add running buffer
  • 05:37 12.3
    Vortex samples & protein marker
  • 05:47 12.4
    Centrifuge samples & protein marker
  • 05:51 12.5
    Loading samples & protein marker
  • 06:07 12.6
    Attach the electrophoresis apparatus to the power supply
  • 06:12 12.7
    Apply the voltage of 80V to the gel, run 15 min
  • 06:18 12.8
    After the dye front has moved into the stacking gel, increase the voltage to 140V and run 50 min
  • Index
  • Notes
  • Comment
  • Fullscreen
Western Blot - 2_Electrophoresis [Video from GeneTex]
Duration: 06:35, Browse: 996, Last Updated: 2018-12-19
Keypoint
  1. 1.
    Leaking test
    add H2O
  2. 2.
    Leaking test
    Confirmed that there is no residual H2O between glass plates
  3. 3.
    Preparation of seperating gel
    <15 kDa         20%
    15-50 kDa       15%
    >50 kDa         12%   
  4. 4.
    Preparation of seperating gel
    .
  5. 5.
    Preparation of stacking gel
    .
  6. 6.
    Insert a clean comb into the stacking gel solution
    .
  7. 7.
    Add running buffer
    .
    • 00:00 1.
      Western Blot - 2_Electrophoresis [Video from GeneTex]
    • 00:19 2.
      Clean the glass plates
    • 00:32 3.
      Assemble the gel apparatus
    • 00:54 4.
      Leaking test
    • 01:03 4.1
      Pour off H2O
    • 01:11 5.
      Preparation of separating gel
    • 01:23 5.1
      Add H2O
    • 01:32 5.2
      Add 1.5M Tris (pH8.8)
    • 01:43 5.3
      Add 30% polyacrylamide
    • 01:54 5.4
      Add 10% SDS
    • 02:08 5.5
      Add 10% ammonium persulfate (APS)
    • 02:22 5.6
      Add TEMED
    • 02:40 5.7
      Add the mixture into the gap between the glass plates
    • 02:51 6.
      Overlay with 75% EtOH
    • 02:57 6.1
      Remove 75% EtOH
    • 02:57 6.2
      Remove 75% EtOH
    • 03:01 7.
      Waiting for 30-60 min
    • 03:05 8.
      Preparation of stacking gel
    • 03:05 8.1
      Add H2O
    • 03:25 8.2
      Add 1M Tris (pH6.8)
    • 03:33 8.3
      Add 30% polyacrylamide
    • 03:45 8.4
      Add 10% SDS
    • 03:55 8.5
      Add 10% ammonium persulfate (APS)
    • 04:07 8.6
      Add TEMED
    • 04:23 8.7
      Add the mixture into the gap between the glass plates
    • 04:31 9.
      Insert a clean comb into the stacking gel solution
    • 04:47 10.
      Waiting for 15-30 min
    • 04:52 11.
      Remove the comb gently
    • 05:09 12.
      Running the gel
    • 05:09 12.1
      Assemble the electrophoresis apparatus
    • 05:29 12.2
      Add running buffer
    • 05:37 12.3
      Vortex samples & protein marker
    • 05:47 12.4
      Centrifuge samples & protein marker
    • 05:51 12.5
      Loading samples & protein marker
    • 06:07 12.6
      Attach the electrophoresis apparatus to the power supply
    • 06:12 12.7
      Apply the voltage of 80V to the gel, run 15 min
    • 06:18 12.8
      After the dye front has moved into the stacking gel, increase the voltage to 140V and run 50 min
    Location
    Folder name
    Western blot (Genetex版本)
    Author
    賴亮全
    Branch
    賴亮全教授
    Created
    2018-12-19 18:19:09
    Last Updated
    2018-12-19 23:32:42
    Duration
    06:35
    Reference
    1