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Viability staining
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Summary
- The supernatant of 3 24-wells plates per donor are put on 1 96-wells plate
Materials & Methods
Materials
Method
Preparations ELISA
- Take 200 μL of supernatant of each well and put it in 96-wells plate
- Divide like shown below, (#plate (24-wells);horizontal lane; vertical lane)
96-wells plate
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01
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02
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03
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04
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05
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06
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07
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08
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09
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10
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11
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12
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A
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1A1
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1A2
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1A3
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1A4
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1A5
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1A6
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2A1
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2A2
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2A3
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2A4
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2A5
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2A6
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B
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1B1
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1B2
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1B3
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1B4
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1B5
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1B6
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2B1
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2B2
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2B3
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2B4
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2B5
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2B6
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C
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1C1
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1C2
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1C3
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1C4
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1C5
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1C6
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2C1
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2C2
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2C3
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2C4
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2C5
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2C6
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D
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1D1
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1D2
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1D3
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1D4
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1D5
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1D6
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2D1
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2D2
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2D3
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2D4
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2D5
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2D6
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E
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3A1
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3A2
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3A3
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3A4
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3A5
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3A6
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F
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3B1
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3B2
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3B3
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3B4
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3B5
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3B6
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G
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3C1
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3C2
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3C3
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3C4
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3C5
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3C6
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H
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3D1
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3D2
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3D3
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3D4
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3D5
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3D6
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- Supernatant is stored @ -20 °C and used for ELISA later on
- Remaining cells are used for viability staining. (See Alamar Blue staining)
Alamar Blue staining
- Remove remaining medium from 24-wells plates
- Wash twice with PBS
- Add per well 475 μL HBSS + 25 μL Alamar blue
- For ~75 wells that's 35.625 mL HBSS + 1875 μL Alamar blue
- Incubate ~30 min. 37 °C
- Measure in Wallace 1420 Victor 2TM, excitation 544 nm emission 390 nm
- This shows the metabolic (mitochondria) activity of cells
Results
- 1 D9 (P21)
- {{todo| 1 D12 (P27)}
- 2 D9 (P21)
- {{todo| 2 D12 (P27)}
- 3 D9 (P21)
- 3 D12 (P27)
Conclusion
Related topics
20.109(S08):Testing_cell_viability_(Day3)
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