Composition of EuroFlow-PID panels

and technical information on reagents

Version 1.2 (14 September 2022)

Published in: J.J.M. van Dongen, M. van der Burg, T. Kalina, M. Perez-Andres, E. Mejstrikova, M. Vlkova,
E. Lopez-Granados, M. Wentink, A.K. Kienzler, J.Philippé, A.E. Sousa, M.C. van Zelm, E. Blanco, A. Orfao.
EuroFlow-based flowcytometric diagnostic screening and classification of primary immunodeficiencies of
the lymphoid system. Frontiers Immunol, in press

©EuroFlow Page 1 of 9
14 September 2022 EuroFlow-PID antibody panels version 1.2

Updated in Version 1.2:

• Correction of small typing errors
• Addition of nuTdT reagent with details to table on page 7
• Addition of cyIgM reagent with details to table on page 7
• Correction of typing errors in Panel 5 composition on page 7

EuroFlow DISCLAIMER

Disclaimer on EuroFlow reference reagents

EuroFlow reference reagents have been obtained from multiple different companies and have
been tested under specific EuroFlow conditions and for usage at specific concentrations in the
EuroFlow antibody combinations, as reported in the EuroFlow documents. The recommended
reagents aim to have comparable results in the (inter-laboratory) standardized EuroFlow staining
procedures. However, EuroFlow cannot be responsible for any change in the manufacturing of
the reagents, such as concentration, purity, dye/protein ratio, or any other factor that may affect
the performance of these products both as single reagents and in antibody combinations. The
companies are responsible for consistency, quality and stability of their products.

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14 September 2022 EuroFlow-PID antibody panels version 1.2

Panel 1. Composition of the 8-color PID Orientation panel and technical

information on reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCH7

CD27 CD45RA CD8 and CD16 and CD4 and CD19 and CD3 CD45
smIgD CD56 smIgM TCRγδ

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
Orientation panel, T cell panel,
CD3 APC SK7 BD Biosciences 345767 2.5
SCID/RTE panel
CD4 PerCPC5.5 SK3 BD Biosciences 332772 Orientation panel 7
CD8 FITC SKI BD Biosciences 345772 Orientation panel 5
CD16 PE 3G8 BD Biosciences 555407 Orientation panel 5
Orientation panel, PreGC8 panel,
CD19 PECy7 J3-119 Beckman Coulter IM3628 PostGC panel, IGH-isotype panel, 5
BM-BCP panel
CD27 BV421 M-T271 BD Biosciences 562513 Orientation panel, PreGC8 panel, 1
PostGC panel, T cell panel
CD45 APCH7 2D1 BD Biosciences 641417 Orientation panel 2
CD45RA BV510 HI100 BD Biosciences 563031 Orientation panel 2.5
CD56 PE C5.9 Cytognos CYT-56PE Orientation panel 5
smIgD FITC IA6-2 BioLegend 348205 Orientation panel 1.25
smIgM PerCPCy5.5 MHM-88 BioLegend 314511 Orientation panel 2
Orientation panel, T cell panel,
TCRγδ PECy7 11F2 BD Biosciences 655410 1
SCID/RTE panel

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Panel 2. Composition of PreGC panel and technical information on reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCAF750

CD27 smIgM CD38 CD5 smIgD CD19 CD21 CD24

Catalogue Application in
Marker Fluorochrome Clone Source µl/test
number EuroFlow-PID panel
CD5 PE UCHT-2 BioLegend 300608 PreGC panel 5
Orientation tube, PreGC panel,
CD19 PECy7 J3-119 Beckman Coulter IM3628 PostGC panel, IGH-isotype panel, 5
BM-BCP panel
PreGC panel, PostGC panel, IGH-
CD21 APC B-ly4 BD Biosciences 559867 10
isotype panel
CD24 APCAF750 ALB9 Beckman Coulter B10738 PreGC panel 5
CD27 BV421 M-T271 BD Biosciences 562513 Orientation tube, PreGC panel, 1
PostGC panel, IGH-isotype panel,
T-cell panel
CD38 FITC HB7 BD Biosciences 340909 PreGC panel 5
smIgD PerCPCy5.5 IA6-2 BioLegend 348208 PreGC panel, PostGC panel, IGH- 1.5
isotype panel
PreGC panel, PostGC panel, IGH-
smIgM BV510 MHM-88 BioLegend 314521 1.3
isotype panel, BM-BCP panel

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Panel 3. Composition of PostGC panel and technical information on reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCA750

smIgE and smIgG and
CD27 smIgM smIgD CD19 CD21 CD38
smIgA smIgA

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
Orientation tube, PreGC panel,
CD19 PECy7 J3-119 Beckman Coulter IM3628 PostGC panel, IGH isotype panel, 5
BM-BCP panel
PreGC panel, PostGC panel, IGH-
CD21 APC B-ly4 BD Biosciences 559867 10
isotype panel
CD27 BV421 M-T271 BD Biosciences 562513 Screening panel, PreGC panel, 1
PostGC panel, IGH-isotype panel,
T cell panel
CD38 APCA750 HB7 BD Biosciences 656646 PostGC panel, IGH-isotype panel 3
smIgA FITC IS11-8E10 Miltenyi 130-093-071 PostGC panel 1
smIgA PE IS11-8E10 Miltenyi 130-093-128 PostGC panel 1
smIgD PerCPCy5.5 IA6-2 BioLegend 348208 PreGC panel, PostGC panel, IGH- 1.5
isotype panel
smIgE FITC polyclonal Life Technologies H15701 PostGC panel 2
smIgG PE G18-145 BD Biosciences 555787 PostGC panel 20
PreGC panel, PostGC panel, IGH-
smIgM BV510 MHM-88 BioLegend 314521 1.3
isotype panel, BM-BCP panel

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Panel 4. Composition of IgH-isotype panel and technical information on

reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCA750

smIgG4 smIgG1
CD27 smIgM and and smIgD CD19 CD21 CD38
smIgG2 smIgG2
smIgG3 smIgA2
CD27 SmIgM and and smIgD CD19 CD21 CD38
SmIgA1 smIgA1

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
Orientation tube, PreGC panel,
CD19 PECy7 J3-119 Beckman Coulter IM3628 PostGC panel, IGH-isotype panel, 5
BM-BCP panel
PreGC panel, PostGC panel, IGH-
CD21 APC B-ly4 BD Biosciences 559867 10
isotype panel
CD27 BV421 M-T271 BD Biosciences 562513 Screening panel, PreGC panel, 1
PostGC panel, IGH-isotype panel,
T cell panel
CD38 APCA750 HB7 BD Biosciences 656646 PostGC panel, IGH-isotype panel 3
smIgA1 FITC SAA1 Cytognos CYT-IGA1F IgH-isotype panel 3
smIgA1 PE SAA1 Cytognos CYT-IGA1PE IgH-isotype panel 3
smIgA2 PE IS11-8E10 Miltenyi 130-093-128 IGH-isotype panel 3
smIgD PerCPCy5.5 IA6-2 BioLegend 348208 PreGC panel, PostGC panel, IGH- 1.5
isotype panel
smIgG1 PE SAG1 Cytognos CYT-IGG1PE IgH-isotype panel 3
smIgG2 FITC SAG2 Cytognos CYT-IGG2F IgH-isotype panel 3
smIgG2 PE SAG2 Cytognos CYT-IGG2PE IgH-isotype panel 3
smIgG3 FITC SAG3 Cytognos CYT-IGG3F IgH-isotype panel 3
smIgG4 FITC SAG4 Cytognos CYT-IGG4PE IgH-isotype panel 3
smIgM BV510 MHM-88 BioLegend 314521 IgH-isotype panel 2

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Panel 5. Composition of BM-BCP panel and technical information on reagents

PacB BV510 BV610 FITC PE PECF594 PerCPCy5.5 PECy7 APC APCC750

CD20 smIgM CD38 nuTdT cyCD79a smIgD cyIgM CD19 CD34 CD10

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
CD10 APCC750 HI10a Cytognos CYT-10AC750 BM-BCP panel 3
CD19 PECy7 J3-119 Beckman Coulter IM3628 Orientation, BM-BCP panel, 5
PreGC panel, PostGC panel, IgH-
isotype panel
CD20 PacB 2H7 Biolegend 302320 BM-BCP panel 1
CD34 APC 8G12 BD Biosciences 345804 BM-BCP panel 5
CD38 BV605 HIT2 Biolegend 303532 BM-BCP panel
cyCD79a PE HM57 Dako R7159 BM-BCP panel 3
smIgD PECF594 IA6-2 BD Biosciences 562540 BM-BCP panel 5
smIgM BV510 MHM-88 BioLegend 314521 BM-BCP panel, PreGC panel, 2
PostGC panel, IgH-isotype panel
cyIgM PerCPCy5.5 MHM-88 BioLegend 314512 BM-BCP panel 2
nuTdT FITC HT6 Dako F7139 BM-BCP panel 10

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Panel 6. Composition of T cell panel and technical information on reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCAF750

CD27 CD4 CD45RO CD197 CD28 TCRγδ CD3 CD8

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
Orientation panel, T cell panel,
CD3 APC SK7 BD Biosciences 345767 2.5
SCID/RTE panel
Orientation panel, T cell panel,
CD4 BV510 OKT4 BioLegend 317443 1.5
SCID/RTE panel
Orientation panel, T cell panel,
CD8 APCAF750 B9.11 Beckman Coulter A94683 1.5
SCID/RTE panel
CD27 BV421 M-T271 BD Biosciences 562513 Orientation panel, PreGC8 panel, 1
PostGC panel, IGH-isotype panel,
T cell panel
CD28 PerCPCy5.5 CD28.2 BioLegend 302921 T cell panel 4
CD45RO FITC UCHL1 Exbio 1F-498-T100 T cell panel, SCID/RTE panel 10
CD197 PE FR 11-11E8 Miltenyi 130-093-621 T cell panel 5
Orientation panel, T cell panel,
TCRγδ PECy7 11F2 BD Biosciences 649806 1
SCID/RTE panel

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Panel 7. Composition of SCID/RTE panel and technical information on reagents

BV421 BV510 FITC PE PerCPCy5.5 PECy7 APC APCAF750

CD62L CD4 CD45RO CD31 HLADR TCRγδ CD3 CD8

Catalogue
Marker Fluorochrome Clone Source Application in EuroFlow panel µl/test
number
Orientation panel, T cell panel,
CD3 APC SK7 BD Biosciences 345767 2.5
SCID/RTE panel
Orientation panel, T cell panel,
CD4 BV510 OKT4 BioLegend 317443 1.5
SCID/RTE panel
Orientation panel, T cell panel,
CD8 APCAF750 B9.11 Beckman Coulter A94683 1.5
SCID/RTE panel
CD31 PE MEM-05 Exbio 1P-273-T100 SCID/RTE panel 5
CD45RO FITC UCHL1 Exbio 1F-498-T100 T-cell panel, SCID/RTE panel 10
CD62L BV421 DREG-56 BioLegend 304827 SCID/RTE panel 2
HLADR PerCPCy5.5 L243 BioLegend 307629 SCID/RTE panel 1.5
Orientation panel, T cell panel,
TCRγδ PECy7 11F2 BD Biosciences 649806 1
SCID/RTE panel

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