Flow cytometry antibodies, kits, reagents

Murine No Compensation Panels

Murine peripheral blood panels

Immunophenotype without Compensation

Immunophenotype without Compensation

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The no compensation panels below have been optimized for murine peripheral blood and secondary lymphoid tissue such as spleen, bone marrow and lymph nodes, to help identify common cell subsets. Useful to create a simple four color panel or a great starting point when designing larger and more complex panels.These panels have been optimized by titrating the antibody to give the best signal resolution.


General Identification Panels
 

T, B and myeloid cells

 

T, B and myeloid cells

Markers

Formats

Catalog #

Dilution

CD3

Pacific Blue

MCA500PB

Neat

CD19

Alexa Fluor 647

MCA1439A647

1:10

CD11b

RPE-Alexa Fluor 750

MCA74P750

1:10

Ly6G

FITC

MCA6077F

1:10

Cells

Surface Markers

T cells

CD3+ CD11b- CD19- Ly6G-

B cells

CD3- CD11b- CD19+ Ly6G-

Monocytes/Macrophages

CD3- CD11b+ CD19- Ly6G-

Granulocytes

CD3- CD11b+ CD19- Ly6G+

Fig 1. General panel. A, Identification of T and B cells. B, Identification of myeloid cells from the CD3- CD19- population. Data acquired on the ZE5 Cell Analyzer.

Fig. 1. General panel. A, identification of T and B cells. B, identification of myeloid cells from the CD3- CD19- population. Data acquired on the ZE5TM Cell Analyzer. 

Isotype controls - Reagents to determine specific binding

 

T, B, NK and myeloid cells

Markers

Formats

Catalog #

Dilution

CD3

Pacific Blue

MCA500PB

Neat

B220

RPE-Alexa Fluor 750

MCA1258P750

1:10

CD49b

FITC

MCA2125F

1:5

CD11b

Alexa Fluor 647

MCA74A647

1:10

Cells

Surface Markers

T cells

CD3+ CD19- CD49- CD11b-

B cells

CD3- CD19+ CD49- CD11b-

NK cells

CD3- CD19- CD49b+ CD11b-/+

Myeloid

CD3- CD19- CD49b- CD11b+

Fig. 1. General panel.

Fig. 2. General panel. A, identification of T and B cells. B, identification of NK cells and myeloid cells using CD3, B220, CD49b and CD11b. Data acquired on the ZE5 Cell Analyzer. 


Lymphocyte Panels
 

T cells

 

T cells

Markers

Formats

Catalog #

Dilution

CD3

Pacific Blue

MCA500PB

Neat

CD8

RPE-Alexa Fluor 750

MCA609P750

1:5

CD4

Alexa Fluor 647

MCA4635A647

1:2

CD45

FITC

MCA1031F

1:5

Cells

Surface Markers

T helper

CD45+ CD3+ CD4+ CD8-

T cytotoxic

CD45+ CD3+ CD8+ CD4-

Fig. 2. T cell panel.

Fig. 3. T cell panel. A, identification of T cells. B, identification of T helper and T cytotoxic cells using CD45, CD3, CD4 and CD8. Data acquired on the ZE5 Cell Analyzer.

Incorporating CD44 and CD62L into this panel will allow identification of naïve, memory and effector T cells for both the helper and cytotoxic populations.

Cells

Surface Markers

Naive T helper cells

CD3+ CD4+ CD62L+ CD44-

Memory T helper cells

CD3+ CD4+ CD62L-CD44+

Effector T helper cells

CD3+ CD4+ CD62L- CD44mid

Naïve T cytotoxic cells

CD3+ CD8+ CD62L+ CD44-

Memory T cytotoxic cells

D3+ CD8+ CD62L- CD44+

Effector T cytotoxic cells

CD3+ CD8+ CD62LCD44mid

Isotype controls - Reagents to determine specific binding

 

Activated T cells

Markers

Formats

Catalog #

Dilution

CD3

Pacific Blue

MCA500PB

Neat

CD4 or CD8

Alexa Fluor 647

MCA4635A647 or MCA609A647

1:2 or 1:5

B220

RPE-Alexa Fluor 750

MCA1258P750

1:10

CD69

FITC

MCA1580F

1:2

Cells

Surface Markers

Resting T helper cells

CD3+ B220- CD4+ CD69-

Activated T helper cells

CD3+ B220- CD4+ CD69+

Resting T cytotoxic cells

CD3+ B220- CD8+ CD69-

Activated T cytotoxic cells

CD3+ B220- CD8+ CD69+

Isotype controls - Reagents to determine specific binding

 

B cells

Markers

Formats

Catalog #

Dilution

CD19

Pacific Blue

MCA1439PB

1:10

B220

RPE-Alexa Fluor 750

MCA1258P750

1:10

IgM

FITC

HCA040F

1:20

CD23

Alexa Fluor 647

MCA2348A647

1:10

Cells

Surface Markers

Immature B cells

CD19+ B220+ IgMCD23-​

Mature B cells

CD19+ B220+ IgM- CD23+​

B cell maturation in the blood and bone marrow can also be defined using IgM and IgD with immature B cells IgM + IgD- and mature B cells IgM-IgD + . Other markers can be used such as CD93 and CD21.

Isotype controls - Reagents to determine specific binding

 

Naïve and memory B cells

Markers

Formats

Catalog #

Dilution

CD19

PB

MCA1439PB

1:10

B220

RPE-Alexa Fluor 750

MCA1258P750

1:10

CD23

Alexa Fluor 647

MCA2348A647

1:10

CD80

FITC

MCA2462F

1:5

Cells

Surface Markers

Naïve B cells

CD19+ B220+ CD23+ CD80-

Memory B cells

CD19+ B220+ CD23- CD80+

The panel above is a simplification of the staining used to identify naïve and memory B cells, as the expression patterns of these surface markers can be variable compared to CD27 + and IgD expression in humans, which are more defined markers for naïve and memory B cells. Whilst this panel gives an indication of the amount of naïve and memory B cells, other markers could be included for more conclusive identification of these cells and other populations such as marginal zone B cells, depending upon the tissue being stained. These markers include CD38, CD62L, CD73, CD21 and PD-L2.


NK cell and Myeloid Panels
 

NK and NKT cells

 

NK and NKT cells

Markers

Formats

Catalog #

Dilution

CD3

Pacific Blue

MCA500PB

Neat

CD335

RPE-Cy7

MCA5938PECy7

Neat

CD45

Alexa Fluor 647

MCA1031A647

1:10

CD122

FITC

MCA2363F

1:10

Cells

Surface Markers

NK precursors

CD45+ CD3- CD122+ CD335-

NK cells

CD45+ CD3- CD122+ CD335+

NKT cells

CD45+ CD3+ CD122+ CD335+

Fig 4. NK, NKT and NK precursor cells

Fig 4. NK, NKT and NK precursor cells. A, identification of NK lineage cells using CD122. B, identification of CD335+ NK and CD335 + CD3 + NKT cells in blood. C, CD122 + CD335- NK cell precursors can also be seen in bone marrow. Data acquired on the ZE5 Cell Analyzer.

Isotype controls - Reagents to determine specific binding

 

Myeloid Cell Panel

Markers

Formats

Catalog #

Dilution

CD11b

Pacific Blue

MCA711PB

1:10

CD11c

RPE-Alexa Fluor 750

MCA1369P750

1:2

Ly6C

Alexa Fluor 647

MCA2389A647

1:10

Ly6G

FITC

MCA6077F

1:10

Cells

Surface Markers

Granulocytes

CD3- CD11b+ CD11c- Ly6G+

Classical Monocytes

Ly6Chi CD11b+ CD11c- Ly6G-

Non-Classical monocytes

Ly6Clo CD11b+ CD11c- Ly6G-

Dendritic cells

CD11c+ CD11b+ Ly6C+/- Ly6G+/-

Fig 5. Myeloid cells

Fig 5. Myeloid cells. A, identification of dendritic cells (DC) and granulocytes. B, monocytes. C, Classical Ly6Chi and non-classical Ly6Clo monocytes. Data acquired on the ZE5 Cell Analyzer.


Although these no compensation panels are not available as pre-mixed four color panels cocktails, they have been optimized on the ZE5 Cell Analyzer and titrated for optimal resolution. To purchase pre-mixed panels for human, mouse, rat or dog cell surface markers, visit our antibody panels and cocktails webpage. If you can’t find the antibody you are interested in, you can search our database of over 3000 flow cytometry tested antibodies. To help you analyze your samples, information on the proportion of these populations in murine blood and tissues 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.