• 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
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Western Blot - 2_Electrophoresis [Video from GeneTex]
長度: 06:35, 瀏覽: 967, 最近修訂: 2018-12-19
重點
  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
    位置
    資料夾名稱
    Western blot (Genetex版本)
    發表人
    賴亮全
    單位
    賴亮全教授
    建立
    2018-12-19 18:19:09
    最近修訂
    2018-12-19 23:32:42
    長度
    06:35
    引用
    1