LNG037 Timeline
Category: Other
Updated: 6 Mar 2020
Contributors
Created byLydia Navarrete-Galvan
Attachments
Periods
T-zero:
1. Aliquot out Tzero samples
2. Split cells into 2 groups
--> EDTA Control (Tube A)
-->No EDTA (Tube B)
3. Put treatment groups into incubatorIncubate Treatment groups for 2 HoursStain T-zero Group
1. AF674, APC/Cy7, AF488
[30 min]T-2 Hours:
1. Add EDTA to all samples
--> Split Tube B (No EDTA) into 2 groups
===> Tube B1 (No wait)
===> Tube B2 (30 min wait)Tube B2
Incubate w/ EDTA
@ room temp - 30 minStain T-2 Hour Group
1. AF674, APC/Cy7, AF488
[30 min]Stain T-2 Hour Tube B2
1. AF674, APC/Cy7, AF488 - 30 min
2. Unlabeled CD107a block - 15 min
3. Three (3) Washes - 45 min
4. PE Stain - 30 min
5. FACS Lyse Fix - 10 min
6. Two (2) Washes - 25 minStain T-zero Group
2. Unlabeled CD107a block
[15 min]Stain T-zero Group
3. Three (3) Washes
[40 min]Stain T-zero Group
4. PE Stain
[30 min]Stain T-zero Group
5. FACS Lyse Fix
[10 min]Stain T-zero Group
6. Two (2) Washes - 25 minT-2 Hours
2. Spin down:
--> Tube A (EDTA Control)
--> Tube B1 (No EDTA, then EDTA)Stain T-2 Hour Group
2. Unlabeled CD107a block
[15 min]Stain T-2 Hour Group
3. Three (3) Washes
[40 min]Stain T-2 Hour Group
4. PE Stain
[30 min]Stain T-2 Hour Group
5. FACS Lyse Fix - 10 minStain T-2 Hour Group
6. Two (2) Washes - 25 min
Comments