Table 3.
Antibody tritation
| Markers | Antibody concentration (μl) | ||||
|---|---|---|---|---|---|
| |
5 |
10 |
20 |
40 |
80 |
| CD45 FITC |
2.0 |
2.0 |
2.0 |
1.9 |
1.9 |
| CD34 FITC |
1.8 |
1.9 |
2.0 |
1.9 |
1.9 |
| CD14 FITC |
1.9 |
1.9 |
1.9 |
1.9 |
1.9 |
| HLADR PE |
0.8 |
0.7 |
0.7 |
0.8 |
0.9 |
| CD73 PE |
1.8 |
1.9 |
2.0 |
2.0 |
2.1 |
| CD83 FITC |
1.7 |
1.8 |
1.8 |
1.9 |
2.0 |
| CD40 PE |
1.5 |
1.7 |
1.6 |
1.7 |
1.8 |
| CD80 FITC |
1.9 |
1.9 |
1.9 |
1.9 |
2.0 |
| CD86 PE |
1.1 |
1.1 |
1.1 |
1.2 |
1.5 |
| HLADR PercPCy5.5 |
1.2 |
1.2 |
1.3 |
1.2 |
1.3 |
|
CD45RA FITC/CD45RO PE/CD3 PercP/CD8 APC |
1.7 |
1.9 |
1.5 |
1.5 |
1.7 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD8 APC |
2.6 |
3.0 |
2.5 |
2.0 |
2.2 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD8 APC |
2.5 |
2.5 |
2.5 |
1.9 |
2.1 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD8 APC |
2.4 |
3.3 |
2.6 |
2.5 |
1.7 |
|
CD45RA FITC/CD45RO PE/CD3 PercP/CD4 APC |
1.8 |
1.7 |
1.4 |
1.3 |
1.4 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD4 APC |
2.2 |
2.1 |
2.2 |
2.2 |
2.1 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD4 APC |
2.5 |
2.4 |
2.4 |
2.2 |
2.1 |
| CD45RA FITC/CD45RO PE/CD3 PercP/CD4 APC |
5.1 |
5.0 |
4.5 |
4.6 |
4.5 |
|
CD3 FITC/CD8 PE/CD4 APC/CD45 PercP |
2.2 |
2.0 |
2.0 |
2.0 |
1.9 |
| CD3 FITC/CD8 PE/CD4 APC/CD45 PercP |
2.6 |
1.8 |
1.8 |
1.2 |
1.2 |
| CD3 FITC/CD8 PE/CD4 APC/CD45 PercP |
4.3 |
4.2 |
4.0 |
4.0 |
4.0 |
| CD3 FITC/CD8 PE/CD4 APC/CD45 PercP |
4.0 |
3.9 |
3.8 |
3.7 |
3.6 |
|
CD3 FITC/CD56-16 PE/CD19 APC/CD45 PercP |
2.1 |
2.4 |
2.4 |
2.3 |
2.1 |
| CD3 FITC/CD56-16 PE/CD19 APC/CD45 PercP |
0.9 |
1.2 |
1.6 |
1.2 |
1.1 |
| CD3 FITC/CD56-16 PE/CD19 APC/CD45 PercP |
2.8 |
1.8 |
3.4 |
2.4 |
1.9 |
| CD3 FITC/CD56-16 PE/CD19 APC/CD45 PercP |
3.1 |
3.3 |
3.7 |
3.7 |
3.7 |
|
Markers |
Antibody concentration (μl) |
||||
| |
2 |
5 |
10 |
20 |
40 |
| CD105 APC |
1.8 |
1.8 |
1.8 |
1.8 |
1.8 |
| CD90 FITC |
1.8 |
1.8 |
1.9 |
1.9 |
1.8 |
|
Markers |
Antibody concentration (μl) |
||||
| |
1 |
2 |
5 |
10 |
20 |
| CD11c APC |
1.6 |
1.4 |
1.3 |
1.4 |
1.4 |
| CD19 APC | 2.3 | 2.8 | 2.9 | 2.7 | 2.4 |
The immunophenotype validation required a first step which is the tritration of each antibody performed by using scalar antibody dilution. The best antibody concentration was that with a higher resolution index. The lowest antibody concentration was chosen when there was an equal resolution index.