Flow cytometry antibodies, kits, reagents

Human No Compensation Panels

Human peripheral blood panels

Immunophenotype without Compensation

Immunophenotype without Compensation

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The no compensation panels below have been optimized for human peripheral blood to help identify common cell subsets. Useful as a simple four color panel or a great starting point when designing larger and more complex panels.


General Identification Panels
 

T, B and myeloid cells

 

T, B and myeloid cells

Markers Formats Catalog # 
CD3 RPE-Alexa Fluor 750 MCA463P750
CD19   Pacific Blue MCA1940PB
CD14 Alexa Fluor 647 MCA1568A647
CD66b FITC MCA216F
Cells Surface Markers
T cells CD3+ CD19- CD14- CD66b-
B cells CD3 -  CD19 +  CD14- CD66b-
Monocytes CD3- CD19- CD14+ CD66b-
Granulocytes CD3- CD19- CD14lo CD66b+

Fig. 1. General panel.

Fig. 1. General panel. A, Identification of lymphoid and B, myeloid cells using CD3, CD19, CD14 and CD66b

Isotype controls - Reagents to determine specific binding

 

T, B and NK cells

Markers Formats Catalog # 
CD3 FITC MCA463F
CD45 Pacific Blue MCA87PB
CD19 RPE-Alexa Fluor 750 MCA1940P750
CD16 / CD56 Alexa Fluor 647 MCA2537A647 / MCA2693A647
Cells Surface Markers
T cells CD3+ CD19- CD45+ CD16- CD56-
B cells CD3- CD19+ CD45+ CD16- CD56-
NK cells CD3- CD19- CD45+ CD16+ CD56+

T Cell Panels
 

Helper and cytotoxic T cells

 

Helper and cytotoxic T cells

Markers Formats Catalog #
CD3 RPE-Alexa Fluor 750 MCA463P750
CD4 Alexa Fluor 647 MCA1267A647
CD45 Pacific Blue MCA87PB
CD8 FITC MCA1226F
Cells Surface Markers
Helper T cells CD45+ CD3+ CD4+ CD8-
Cytotoxic T cells CD45+ CD3+ CD4- CD8+
Isotype controls - Reagents to determine specific binding

 

Activated T cells

Markers Formats Catalog #
CD3 Alexa Fluor 647 MCA463A647
CD4 or CD8 Pacific Blue MCA1267PB/MCA1226PB
CD38 RPE-Alexa Fluor 750 MCA1019P750
HLA-DR FITC MCA71F
Cells Surface Markers
Resting T helper cells CD3+ CD4+ CD38- or HLA-DR-
Activated T helper cells CD3+ CD4+ CD38+ or HLA-DR+
Resting T cytotoxic cells CD3+ CD8+ CD38- or HLA-DR-
Activated T cytotoxic cells CD3+ CD8+ CD38+ or HLA-DR+

Fig. 2. Identification of activated CD8 T cells.

Fig. 2. Identification of activated CD8 T cells. A, CD3 and CD8 staining. B, HLA-DR and CD38 shows activated cells in the top right quadrant.

Isotype controls - Reagents to determine specific binding

 

Memory and naïve T cells

Markers Formats Catalog #
CD3 RPE-Alexa Fluor 750 MCA463P750
CD4 / CD8 Alexa Fluor 647 MCA1267A647 or MCA1226A647
CD45RO FITC       MCA461F
CD45RA Pacific Blue MCA88PB
Cells Surface Markers
Naïve T helper cells CD3+ CD4+ CD45RA+ CD45RO-
Naïve T cytotoxic cells    CD3+ CD8+ CD45RA+ CD45RO-
Memory T helper cells   CD3+ CD4+ CD45RA- CD45RO+
Memory T cytotoxic cells CD3+ CD8+ CD45RA- CD45RO+

Fig. 3. Identification of memory and naïve CD4 T cells.

Fig. 3. Identification of memory and naïve CD4 T cells. A, CD3 and CD4 staining. B, CD45RA shows naïve cells in the top left and CD45RO shows memory cells in the bottom right quadrants. 

Incorporating CD27 or CD62L into this panel will allow the identification of effector T cells.

Cells Surface Markers
Memory T helper cells   CD3+ CD4+ CD45RO+ CD27 and/or CD62L+
Effector T helper cells    CD3+ CD4+ CD45RO- CD27 and/or CD62L-
Memory T cytotoxic cells CD3+ CD8+ CD45RO+ CD27 and/or CD62L+
Effector T cytotoxic cells CD3+ CD8+ CD45RO- CD27 and/or CD62L-
Isotype controls - Reagents to determine specific binding

 

Treg cells (when permeabilizing is not required)

Markers Formats Catalog #
CD3 RPE-Alexa Fluor 750 MCA463P750
CD4 PB Pacific Blue MCA1267PB
CD25 FITC MCA2127F
CD127 Alexa Fluor 647 HCA145A647
Cells Surface Markers
Treg cells CD3+ CD4+ CD127lo CD25hi

Fig. 4. Identification of regulatory T cells. 

Fig. 4. Identification of regulatory T cells. A, CD3 and CD4 staining. B, CD127 and CD25 staining reveals the Treg population.


B Cell Panels
 

Memory and naïve B cells

 

Memory and naïve B cells

Markers Formats Catalog #
CD19 RPE-Alexa Fluor 750 MCA1940P750
CD20 Pacific Blue MCA1710PB
IgD FITC STAR143F
CD27 Alexa Fluor 647 MCA755A647
Cells Surface Markers
Memory B cells CD19+ CD20+ CD27+ IgD-
Naïve B cells CD19+ CD20+ CD27- IgD+

Fig. 5. Naïve and memory B cells.

Fig. 5. Naïve and memory B cells. A, forward and side scatter. B, identification of lymphocyte population. C, naïve B cells can be seen in the bottom right and memory B cells in the top left quadrants.

Isotype controls - Reagents to determine specific binding

 

Transitional B cells and plasmablasts

Markers Formats Catalog #
CD19 Alexa Fluor 647 MCA1940A647
CD20 Pacific Blue MCA1710PB
CD24 FITC MCA1379F
CD38 RPE-Alexa Fluor 750 MCA1019P750
Cells Surface Markers
Transitional B cells CD19+ CD20+ CD24+ CD38+
Plasmablasts CD19+ CD20+ CD24- CD38+
Isotype controls - Reagents to determine specific binding

 

Class switched memory B cells

Markers Formats Catalog #
CD19 RPE-Alexa Fluor 750 MCA1940P750
CD20 Pacific Blue MCA1710PB
CD27 Alexa Fluor 647 MCA755A647
IgG FITC 204002F
Cells Surface Markers
Non-class-switched memory B cells CD19+ CD20+ CD27+ IgG-
Class-switched memory B cells CD19+ CD20+ CD27+ IgG+

NK Cells and Myeloid Panels
 

NK and NKT cells

 

NK and NKT cells

Markers Formats Catalog #
CD3 RPE-Alexa Fluor 750 MCA463P750
CD45 Pacific Blue MCA87PB
CD56 Alexa Fluor 647 MCA2693A647
CD335 FITC MCA4646F
Cells Surface Markers
T cells CD3+, CD45+, CD56-, CD335-
NK cells CD3-, CD45+, CD56+, CD335+
NKT cells CD3+, CD45+, CD56+

Fig. 6. Identification of NK and NKT cells

Fig. 6. Identification of NK and NKT cells. A, CD45 and CD3 staining. B, gated on the Non T, CD335 and CD56 staining reveals the NK population. C, gated on the T cell population reveals the NKT cells.

Isotype controls - Reagents to determine specific binding

 

Myeloid cells

Markers Formats Catalog #
CD14 RPE-Alexa Fluor 750 MCA1568P750
CD66b FITC MCA216F
CD45 Pacific Blue MCA87PB
CD16 Alexa Fluor 647 MCA2537A647
Cells Surface Markers
Monocytes CD45+ CD16+ CD14+ CD66b-
Granulocytes CD45+ CD16+ CD14lo CD66b+

These no compensation panels are not available as pre-mixed four color panel cocktails and therefore may need to be optimized. Pre-optimized panels for human, mouse, rat or dog cell surface markers are available on our antibody panels and cocktails webpage. If you can’t find the antibody you are interested in, search our database of over 3000 flow cytometry tested antibodies.

To help you analyze your samples, information on the proportion of these populations in human blood can be found on our cell frequency page.

When performing multicolor flow cytometry, you should consider careful sample preparation, Fc blocking, inclusion of a live/dead dye, doublet exclusion and using the appropriate controls. For more information, watch our flow cytometry webinar Optimize your Flow Cytometry or visit our application resources page.