Checking Cells
Before collecting the cells, the cells should be checked microscoptically to ensure they are healthy and growing as expected.
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00:18
1.
Western Blot - 1_Sample preparation [Video from GeneTex]
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00:11
2.
Checking cells
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00:12
3.
Remove medium
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00:16
4.
Wash cells
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00:13
5.
Lyse cells
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00:10
6.
Scrape cells
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00:09
7.
Collect cells
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00:06
8.
Vortex
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00:02
9.
Centrifuge
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00:12
9.1
Balance the tubes
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00:11
10.
Collect supernatant
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00:17
11.
Denature
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00:09
11.1
Boil the mixture at 95-100 C for 5 min
- Index
- Notes
- Comment
- Fullscreen
Western Blot - 1_Sample preparation [Video from GeneTex]
Duration: 02:33, Browse: 833, Last Updated: 2018-12-19
Play Video: http://llai.cm.ntu.edu.tw/media/169
Keypoint
- 1.
- 2.RIPA bufferRIPA bufferTris-HCl (pH 7.6) 25mMNaCl 150 mMTriton X-100 1%Na-deoxychola 1%SDS 1%Store at 4CAdd the protease inhibitors immediately prior use
- 3.Centrifuge TipUse the right size of tubes for the right centrifuge. If tube size is too small, you need to add adaptor.
- 4.Centrifuge20,000 g at 4C for 30 min
- 5.Denature6X SDS Sample Buffer:Tris-HCl (pH6.8) 315.5mM
Glycerol 50%
SDS 0.1 g/ml
bromophenol blue 0.05%
2-b mercaptoethanol 10%
-
00:18
1.
Western Blot - 1_Sample preparation [Video from GeneTex]
-
00:11
2.
Checking cells
-
00:12
3.
Remove medium
-
00:16
4.
Wash cells
-
00:13
5.
Lyse cells
-
00:10
6.
Scrape cells
-
00:09
7.
Collect cells
-
00:06
8.
Vortex
-
00:02
9.
Centrifuge
-
00:12
9.1
Balance the tubes
-
00:11
10.
Collect supernatant
-
00:17
11.
Denature
-
00:09
11.1
Boil the mixture at 95-100 C for 5 min
- Location
-
- Folder name
- Western blot (Genetex版本)
- Author
- 賴亮全
- Branch
- 賴亮全教授
- Created
- 2018-12-19 18:18:30
- Last Updated
- 2018-12-19 22:33:54
- Browse
- 833
- Duration
- 02:33
- Reference
- 1