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June 30, 2022
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LNG037 Timeline
Created by
Lydia Navarrete-Galvan
⟶ Updated 6 Mar 2020 ⟶
List of edits
Comments
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 incubator
Incubate Treatment groups for 2 Hours
Stain 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 min
Stain 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 min
Stain 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 min
T-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 min
Stain T-2 Hour Group 6. Two (2) Washes - 25 min
