Key Extracellular Treg Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD3 | OKT3 | Identification of T Cells |
PacBlue |
||
 | UCHT1 | Identification of T Cells |
PacBlue |
FITC PE/Dazzle594 PE/Cy5 |
APC |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
|
CD25 | BC96 | Tregs express high levels of CD25 |
PacBlue BV785 |
FITC |
APC |
 | M-A251 |  | PacBlue (2380645, 2380650)BV421 (2380565, 2380570)BV510 (2380595, 2380600) |
FITC |
APC |
CD127 | A019D5 | Inversly correlates with FoxP3 expression. Useful for T-reg enrichment/isolation. |
PacBlue BV570 |
FITC |
APC (2356575, 2356580)AF647 (2356585, 2356590) |
Key Intracellular Treg Markers
Other Important Treg Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD6 | BL-CD6 | CD6 low expression may be useful in combination with CD127 for Treg enrichment | FITC (2169520)PE (2169530) |
||
CD39 | A1 | Identifies a subset of Tregs. May be important for Treg function | BV421 (2241065, 2241070)BV510 (2241095) |
FITC |
APC (2241045, 2241050) |
CD45RA | HI100 | Useful to distinguish nTreg versus iTreg |
PacBlue |
FITC PE/Dazzle594 PE/Cy5 |
APC (2120555, 2120560)AF700 (2120595, 2120600)APC/Cy7 (2120635, 2120640) |
CD45RO | UCHL1 | T-cell memory/differentiation marker |
PacBlue BV711 |
FITC PE/Cy5 |
APC (2121050)AF700 (2121085, 2121090)APC/Cy7 (2121135, 2121140) |
CD137 | 4B4-1 | Activation marker. Recommended to be used in combination with CD154 | BV421 (2149095, 2149100) |
PerCP/Cy5.5 |
APC (2149045, 2149050)AF700 (2149080) |
CD154 | 24-31 | Activation marker. Recommended to be used in combination with CD137 |
PacBlue BV605 |
APC (2154045, 2154050)AF647 (2154090)APC/Cy7 (2154105, 2154110) |
|
CD183 (CXCR3) |
G025H7 | Possible marker of activated Treg |
PacBlue BV650 |
FITC |
APC (2368535, 2368540)AF647 (2368555, 2368560)APC/Cy7 (2368605, 2368610) |
CD184 (CXCR4) |
12G5 | Involved in Treg migration | BV421 (2132585, 2132590)BV605 (2132605, 2132610) |
PerCP/Cy5.5 |
APC (2132545, 2132550) |
CD194 (CCR4) |
L291H4 | Involved in Treg trafficking; expressed on tumor infiltrating T cells | BV421 (2397065, 2397070)BV510 (2397075, 2397080)BV605 (2397085, 2397090) |
PerCP/Cy5.5 |
APC (2397035, 2397040)AF647 (2397015, 2397020) |
 | TG6/CCR4 |  | PerCP/Cy5.5 (2277015) |
AF647 (2277005) |
|
CD278 (ICOS) |
C398.4A | May identify a Treg subset that expresses higher levels of IL-10 | PacBlue (2167605, 2167610)BV421 (2167615, 2167620)BV510 (2167625) |
FITC |
APC (2167545, 2167550)AF647 (2167575, 2167580) |
CD279 (PD-1) |
EH12.2H7 | May be expessed by tumor infiltrating T-reg |
PacBlue |
FITC |
APC (2249535, 2249540)AF647 (2249550)APC/Cy7 (2249605, 2249610) |
Galectin-9 | 9M1-3 | Reported to increase iTreg stability | FITC (2344555, 2344560)PerCP/Cy5.5 (2344545, 2344550)PE (2344525, 2344530) |
APC (2344535, 2344540) |
|
GARP (LRRC32) |
7B11 | May be useful for the identification of antigen-specific T-regs | BV421 (2362545) |
PE (2362515, 2362520)PE/Cy7 (2362535, 2362540) |
APC (2362525, 2362530 |
HLA-DR | L243 | Identifies a subset of Treg |
PacBlue BV570 |
FITC |
APC |
LAP (TGF-β1) |
TW4-2F8 | May be useful for the identification of antigen-specific T-regs | BV421 (2348065) |
FITC PE/Cy7 |
APC (2348035, 2348040) |
Perforin | B-D48 | Important for Treg functions | PacBlue (2366525)BV421 (2366535) |
Key Intracellular Markers
Th1 Key Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
|
CD183 (CXCR3) |
G025H7 | Highly expressed on Th1 cells; trafficking |
PacBlue BV650 |
FITC |
APC (2368535, 2368540)AF647 (2368555, 2368560)APC/Cy7 (2368605, 2368610) |
Key Th1 Intracellular Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
IFN-γ | 4S.B3 | Th1 signature cytokine |
PacBlue |
FITC |
|
IFN-γ | B27 | Â | FITC (3132520)PE (3132530, 3132535)PE/Cy7 (3132585, 3132590) |
APC (3132550)AF700 (3132575, 3132580) |
|
LT-α (TNF-β) |
359-81-11 | Cytokine expressed by Th1 cells | PE (3115525) |
||
TNF-α | Mab11 | Cytokine expressed by Th1 cells |
PacBlue BV711 |
FITC |
APC (3114560)AF647 (3114580)AF700 (3114635, 3114640) |
T-bet | 4B10 | Key Th1 transcription factor |
PacBlue BV711 |
FITC |
APC (3824065, 3824070)AF647 (3824015, 3824020) |
Th2 Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
|
CD194 (CCR4) |
L291H4 | Th2 homing | BV421 (2397065, 2397070)BV510 (2397075, 2397080)BV605 (2397085, 2397090) |
PerCP/Cy5.5 |
APC (2397035, 2397040)AF647 (2397015, 2397020) |
CD294 (CRTH2) |
BM16 | Th2 marker. May be important in inflammation. | BV421 (2350555, 2350560) |
FITC |
APC (2350545, 2350550)AF647 (2350515, 2350520)APC/Cy7 (2350565, 2350570) |
Intracellular Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
IL-4 | 8D4-8 | Signature cytokine for Th2 cells | AF488 (3103550)PE (3103515, 3103520) |
APC (3103565, 3103570)AF647 (3103560) |
|
 | MP4-25D2 |  | BV421 (3104125)BV605 (3104135, 3104140) |
FITC PE |
APC (3104055, 3104060)AF647 (3104090) |
IL-5 | TRFK5 | Expressed on a subset of Th2 cells | BV421 (3121555(50ug)) |
PE (3121515, 3121520) |
APC (3121525, 3121530) |
IL-13 | JES10-5A2 | Cytokine expressed by Th2 cells | PerCP/Cy5.5 (3109555)PE (3109515) |
APC (3109535) |
|
GATA3 | 16E10A23 | Key transcription factor for Th2 cells | BV421 (3869065, 3869070) |
AF488 (3869035, 3869040)PerCP/Cy5.5 (3869055, 3869060)PE (3869020) |
APC (3869025, 3869030)AF647 (3869045, 3869050) |
Th9 Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
Intracellular Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
IL-9 | MH9A4 | Signature cytokine for Th9 cells | PE (3138020, 3138025) |
AF647 (3138035, 3138040) |
|
BATF | 9B5A13 | Transcription factor expressed by Th9 cells | PE (3874015, 3874020) |
||
IRF4 | IRF4.3E4 | Transcription factor expressed by Th9 cells | AF488 (3832025, 3832030)PE (3832015, 3832020) |
AF647 (3832035, 3832040) |
|
SPI1 (PU.1) |
7C6B05 | Transcription factor expressed by Th9 cells | AF647 (3890015, 3890020) |
Th17 Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
|
CD196 (CCR6) |
G034E3 | Â |
BV421 |
FITC |
APC (2367075, 2367080)AF647 (2367015, 2367020) |
CD39 | A1 | Identifies a subset of Th17 cells | BV421 (2241065, 2241070)BV510 (2241095) |
FITC |
APC (2241045, 2241050) |
CD73 | AD2 | Expressed by a subset of Th cells | PacBlue (Biotin only)BV421 (2320085) |
||
CD161 | HP-3G10 | Expressed on most Th17 cells |
PacBlue |
FITC |
APC (2299555, 2299560)AF647 (2299545, 2299550)APC/Cy7 (2299635, 2299640)Â |
Intracellular Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
IL-17A | BL168 | Signature Th17 cytokine |
PacBlue |
FITC PE/Dazzle594 PE/Cy7 |
APC |
IL-21 | 3A3-N2 | Â | PE (3165015, 3165020) |
APC (3165035, 3165040)AF647 (3165025, 3165030) |
|
TNF-α | Mab11 | Cytokine expressed by Th1 cells |
PacBlue BV711 |
FITC3114530) |
Th22 Cells
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
CD4 | RPA-T4 | Identification of CD4+ helper T cells |
PacBlue BV650 |
FITC |
|
CD194 (CCR4) |
L291H4 | Homing receptor expressed on Th22 cells | BV421 (2397065, 2397070)BV510 (2397075, 2397080)BV605 (2397085, 2397090) |
PerCP/Cy5.5 |
APC (2397035, 2397040)AF647 (2397015, 2397020) |
CD196 (CCR6) |
G034E3 | Homing receptor expressed on Th22 cells |
BV421 |
FITC |
APC (2367075, 2367080)AF647 (2367015, 2367020) |
CCR10 | 6588-5 | Homing receptor expressed on Th22 cells | PE (2307515, 2307520) |
APC (2307525, 2307530) |
Intracellular Markers
Marker | Clone | Purpose | Violet Laser | Blue Laser | Red Laser |
IL-22 | BG/IL-22 | PE (3176515) |
T-Cell Panels
These panels are based on panels that have been published. Additional markers may be added for more information. The first panel is completely extracellular and is therefore suitable for cell sorting and analysis applications. The second one contains FoxP3, an intracellular marker, and is therefore only suitable for the analysis of Tregs. We've designed these panels, based on published panels and included the references below.
Specificity | Clone | Fluorochrome | Purpose | Cat. No. | Name |
CD25 | M-A251 | PE | T cell activation/Treg ID | 2380515 | PE anti-Human CD25 |
CD4 | SK3 | PerCP-Cy5.5 | T cell subset | 2323035 | PerCP/Cy5.5 anti-human CD4 |
CD194 (CCR4) | L291H4 | PE/Cy7 | T cell trafficking | 2397045 | PE/Cy7 anti-human CD194 (CCR4) |
CD127 (IL-7Rα) | A019D5 | Alexa Fluor® 647 | Low expession is correlated with Treg | 2356585 | Alexa Fluor® 647 anti-human CD127 (IL-7Rα) |
CD45RO | UCHL1 | APC/Cy7 | Maturation/Differentiation | 2121135 | APC/Cy7 anti-human CD45RO |
CD3 | UCHL1 | Pacific Blueâ„¢ | T cell marker | 2102090 | Pacific Blueâ„¢ anti-human CD3 |
HLA-DR | LN3 | FITC | Treg Subset | 2235025 | FITC anti-human HLA-DR |
Specificity | Clone | Fluorochrome | Purpose | Cat. No. | Name |
CD3 | UCHT1 | FITC | T cell marker | 2102030 | FITC anti-human CD3 |
CD4 | SK3 | PerCP-Cy5.5 | 2323035 | PerCP/Cy5.5 anti-human CD4 | |
CD45RO | UCHL1 | Brilliant Violet 421â„¢ | Maturation Differentiation | 2121115 | Brilliant Violet 421â„¢ anti-human CD45RO |
CD25 | M-A251 | PE/Dazzleâ„¢ 594 | T cell activation/Treg ID | 280625 | PE/Dazzleâ„¢ 594 anti-human CD25 |
FOXP3 | 206D | PE | Treg Subsets | 2200535 | PE anti-human FOXP3 |
CD39 | A1 | PE-Cy7 | 2241055 | PE/Cy7 anti-human CD39 | |
CD49d | 9F10 | PE-Cy5 | 2121530 | PE/Cy5 anti-human CD49d |
References: http://iti.stanford.edu/himc/flow-cytometry.html
Murdoch, David M., Janet S. Staats, and Kent J. Weinhold. "OMIPâ€006: Phenotypic subset analysis of human T regulatory cells via polychromatic flow cytometry." Cytometry Part A 81.4 (2012): 281-283.