Migration Assay
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00:05
1.
Migration Assay
-
00:24
2.
Put migration chambers i 24-well plate
-
00:18
3.
Add 100ul serum-free medium in the chamber
-
00:23
4.
Add 200ul cell (2.5 x 10^5/ml in serum-free medium) in the chamber
-
00:34
5.
Add 750ul culture medium (WITH serum) in lower chamber
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00:04
5.1
Incubate at 37C for 12-16 hours
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00:11
6.
Remove medium from the chamber
-
00:07
7.
Wash twice in PBS
-
00:25
8.
Fix cells by formaldehyde (3.7% in PBS)
-
00:03
8.1
Fix cells at room temp. for 2 min
-
00:34
9.
Remove formaldehyde and wash twice by PBS
-
00:21
10.
Permeabilize cells by 100% methanol
-
00:05
10.1
Permeabilization at room temp for 20 min
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00:27
11.
Remove methanol and wash twice by PBS
-
00:34
12.
Giemsa stain
-
00:24
13.
Remove Giemsa stain and wash twice by PBS
-
00:15
14.
Scrape off non-migrated cells with cotton swabs
-
00:14
15.
Count migrated cells under a light microscope
Play Video: http://llai.cm.ntu.edu.tw/media/402
( http://www.abnova.com ) - The migration assay (also known as the Boyden Chamber Assay) is a commonly used test to study the migratory response of endothelial cells. During this assay, cells are placed on the upper layer of a chamber and a solution containing the test agent is placed below the chamber.
-
00:05
1.
Migration Assay
-
00:24
2.
Put migration chambers i 24-well plate
-
00:18
3.
Add 100ul serum-free medium in the chamber
-
00:23
4.
Add 200ul cell (2.5 x 10^5/ml in serum-free medium) in the chamber
-
00:34
5.
Add 750ul culture medium (WITH serum) in lower chamber
-
00:04
5.1
Incubate at 37C for 12-16 hours
-
00:11
6.
Remove medium from the chamber
-
00:07
7.
Wash twice in PBS
-
00:25
8.
Fix cells by formaldehyde (3.7% in PBS)
-
00:03
8.1
Fix cells at room temp. for 2 min
-
00:34
9.
Remove formaldehyde and wash twice by PBS
-
00:21
10.
Permeabilize cells by 100% methanol
-
00:05
10.1
Permeabilization at room temp for 20 min
-
00:27
11.
Remove methanol and wash twice by PBS
-
00:34
12.
Giemsa stain
-
00:24
13.
Remove Giemsa stain and wash twice by PBS
-
00:15
14.
Scrape off non-migrated cells with cotton swabs
-
00:14
15.
Count migrated cells under a light microscope
- Location
-
- Folder name
- migration assay (Abnova版本; 伊純)
- Author
- 鄭伊純
- Branch
- 賴亮全教授
- Created
- 2019-01-07 08:16:33
- Last Updated
- 2019-01-07 08:16:34
- Browse
- 1,280
- Duration
- 05:36
- Source
- https://youtu.be/6SON7VAA5-k