PROTOCOL FOR ESTIMATING THE NUMBER OF CD38 MOLECULES ON THE CD8+T LYMPHOCYT

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Tony Rook
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PROTOCOL FOR ESTIMATING THE NUMBER OF CD38 MOLECULES ON THE CD8+T LYMPHOCYT

Please find the link for the following protocol:

http://aactg.s-3.com/pub/download/imm/imcd38.pdf

PROTOCOL FOR ESTIMATING THE NUMBER OF CD38 MOLECULES ON THE CD8+T LYMPHOCYTES OF HIV-INFECTED INDIVIDUALS

1. MATERIALS
2. RECOMMENDATION OF VENDOR FOR PE-CD38 AND PE-CD4
3. VALIDATION OF LOGARITHMIC AMPLIFIER LINEARITY AND SENSITIVITY
3.1. INTRODUCTION
3.2. PREPARATION OF RAINBOW BEADS
3.3. PMT VOLTAGES
3.4. ANALYSIS OF INSTRUMENT PERFORMANCE
3.5. RAINBOW BEAD SPREADSHEET
4. CONSERVATION OF THE LEVEL OF CD4 ANTIGEN EXPRESSION ON CD4+
LYMPHOCYTES AND ITS USE AS A BIOLOGIC STANDARD FOR FLOW CYTOMETER
INSTRUMENT CHARACTERIZATION
5. SAMPLE PREPARATION METHODS 1 AND 2
5.1. METHOD 1: HEALTHY CONTROLS FOR INSTRUMENT CHARACTERIZATION
5.2. METHOD 2: PATIENT SAMPLES FOR CD38 QUANTITATION
5.3. METHODS 1 & 2: COMPENSATION TUBES
5.4. OTHER TUBES
6. DAILY INSTRUMENT SETUP
6.1. INTRODUCTION
6.2. SET PMT VOLTAGES TO PLACE CONTROL BEAD (M6) IN THE TARGET CHANNEL
6.3. SET INSTRUMENT COMPENSATION
6.4. CAUTION
6.5. REPEAT BEFORE EACH RUN
7. CHARACTERIZATION OF INSTRUMENT FOR NUMBER OF MOLECULES OF PE PER RFI
CHANNEL
7.1. OVERVIEW
7.2. MAB PANEL FOR PE-CD4 MEDIAN RFI
7.3. SAMPLE PREPARATION
7.4. SAMPLE ACQUISITION
7.5. DATA ANALYSIS
7.6. HEALTHY CONTROL SPREADSHEET
8. DETERMINATION OF THE NUMBER OF CD38 MOLECULES PER CD8+ CELL
8.1. MAB PANEL FOR PE-CD38 MEDIAN RFI ON CD8
8.2. SAMPLE PREPARATION
8.3. SAMPLE ACQUISITION
8.4. DATA ANALYSIS
8.5. CD38 SPREADSHEET
9. CLINICAL USE OF THE NUMBER OF CD38 MOLECULES PER CD8+ CELL (CD38 ON CD8)