New Reagents for Cancer Research: TCL1, ZAP-70, SLP-76, BAMBI, B7-H1
T cell leukemia-1 (TCL1) is a 14 kDa protein with a suggested role in intracellular regulation of T cell signalling. TCL1 was identified as the oncogene located at the 14q32.1 chromosome breakpoint region in T-cell prolymphocytic leukemia (T-PLL). In T-PLL, TCL1 is
overexpressed as a result of an inversion or a reciprocal translocation, by juxtaposition to the T-cell receptor promoter/enhancer elements. TCL1 binds to the pleckstrin homology domain of Akt (protein kinase B) family proteins, which facilitates Akt dimerization
and activity. By increasing Akt activity TCL1 may enhance the serine/threonine phosphorylation of major Akt signaling substrates, such as Ikk complex, mTOR, BAD, p70S6 kinase, FOXO transcription factors, and GSK3b. These substrates regulate cellular
differentiation, growth, survival, and metabolism.
Besides its tumorigenic role in T-PLL, TCL1 is normally expressed in the CD3-CD4-CD8- subset of thymic precursors in the T cell lineage, the plasmacytoid subset of dendritic cells, stimulated (not resting) mature T cells, and B cells up the germinal center stage of
maturation. TCL1 is inappropriately expressed by chromosome rearrangements that lead to pre-malignant clonal T cell expansions and mature T cell tumors. TCL1 shows a regulated expression pattern in chronic lymphocytic leukemia (CLL). The anti-human TCL-1
antibody, clone eBio1-21, has been reported for use in immunohistochemistry (paraffin, formalin), western blotting, and intracellular staining for flow cytometric analysis (ICFC).
References
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Herling M, Patel KA, Khalili J, Schlette E, Kobayashi R, Medeiros LJ, Jones D. TCL1 shows a regulated expression pattern in chronic lymphocytic leukemia that correlates with molecular subtypes and proliferative state. Leukemia. 2006 Feb;20(2):280-5. [IHC/formalin, Western blotting, PubMed]
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Hoyer KK, Herling M, Bagrintseva K, Dawson DW, French SW, Renard M, Weinger JG, Jones D, Teitell MA. T cell leukemia-1 modulates TCR signal strength and IFN-gamma levels through phosphatidylinositol 3-kinase and protein kinase C pathway activation. J Immunol. 2005 Jul 15;175(2):864-73. [IHC/formalin, Western blotting, PubMed]
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Herling M, Teitell MA, Shen RR, Medeiros LJ, Jones D. TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin. Blood. 2003 Jun 15;101(12):5007-9. Epub 2003 Feb 6. [Western blotting, ICFC, IHC/paraffin, PubMed]
ZAP-70 is a cytosolic protein tyrosine kinase and a member of the Syk family of proteins.
It is expressed in T and NK cells and is required for TCR signaling and development.
Following activation, ZAP-70 is phosphorylated on several tyrosine residues. Mutations in
ZAP-70 have been shown to result in a form of Severe Combined Immunodeficiency
Syndrome (SCID) in humans. While ZAP-70 is normally expressed in T and NK cells,
several recent studies have also shown high correlation of ZAP-70 positive expression
with mutated IgVH expression in B-chronic lymphocytic leukemia (CCL).
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Human/Mouse ZAP-70 (1E7.2):
Staining of fixed/permeabilized human Jurkat T cells
with PE-Mouse IgG1, K Isotype Control (open histogram)
or PE-anti-human/mouse ZAP-70 (colored
histogram).
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SLP-76 is a cytosolic adaptor protein expressed in several cell types of hematopoietic
origin. SLP-76 is required for coupling receptor ligation with more distal signaling events
in immature thymocytes, mature T cells, platelets, and mast cells. SLP-76 was originally
identified as a substrate of the ZAP-70 protein tyrosine kinase following T cell receptor
(TCR) ligation in the leukemic T cell line Jurkat. Studies using SLP-76 deficient T cell lines
or mice have provided strong evidence that SLP-76 plays a positive role in promoting T
cell development and activation.
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Mouse SLP-76 (MS76):
CD3 vs. SLP-76
Staining of fixed/permeabilized
mouse splenocytes with APC anti-mouse CD3
(FL4) and FITC anti-mouse SLP-76 (FL1).
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Annexins are a family of calcium-dependent phospholipid-binding proteins abundant in the eukaryotic kingdom. Annexin V's preferential binding partner is phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet or cytosol-facing part of the plasma membrane. Upon initiation of apoptosis, PS loses its asymmetric distribution in the phospholipid bilayer and is translocated to the extracellular membrane leaflet where it identifies cells as targets of phagocytosis. Once on the outer surface of the membrane, PS can be detected by a fluorescently labeled annexin V in a Ca2+ -dependent manner.
In early stage apoptosis, the plasma membrane excludes viability dyes such as propidium iodide and 7-AAD, so these cells will only stain with Annexin V and serves as an early marker of apoptosis. However, in late stage apoptosis, the cell membrane loses integrity allowing Annexin V to access PS in the interior of the cell, as well. A viability dye such as 7-AAD or propidium iodide (PI) can be used to resolve these late stage apoptotic and necrotic cells (Annexin V and 7-AAD or PI double positive) from the early stage apoptotic cells (Annexin V positive, 7-AAD or PI negative).
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Jurkat cells were either untreated (left) or treated for 4 hours with 10µM camptothecin (right) and then stained with the Annexin V-APC Apoptosis Detection Kit and Propidium Iodide Staining Solution (PI) (cat. 00-6990). The Annexin V-positive, PI-negative population contains early stage apoptotic cells.
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Table 1: Human Cancer Markers
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Human Cancer Markers
|
| Human |
Clone |
SKU |
Applications |
Formats |
|
|
|
|
|
|
| ABCG2/Bcrp1
|
5D3
|
8888
|
FC
|
Purified, FG Purified, Biotin, PE
|
| B7-H1
|
MIH1
|
5983
|
FC, BA, IHC/F
|
Purified, FG Purified, Biotin, PE, PE-Cy7, APC
|
| BAMBI
|
4E8
|
5826
|
FC
|
Purified, Biotin, PE
|
| Bcl-xL
|
2H12
|
6994
|
WB
|
Purified
|
| BR3
|
8A7
|
9117
|
FC, BA, WB, IHC
|
Purified, FG Purified, FITC, PE
|
| E-Cadherin
|
DECMA-1
|
3249
|
FC, ELISA, FA, IP, IHC
|
Purified, Alexa Fluor® 647
|
| EpCAM
|
1B7
|
9326
|
FC, WB, IP, IHC (Paraffin)
|
Purified, PE, Alexa Fluor® 647
|
| HLA-G
|
87G
|
9957
|
FC, IHC/F
|
Purified, PE
|
| FREB
|
N28.1
|
5847
|
ICFC, IHC
|
Purified, Alexa Fluor® 647
|
| RASSF1A
|
eB114-10H1
|
6888
|
WB, IP
|
Purified
|
| SLP-76
|
HS76
|
7769
|
ICFC, WB
|
Purified, FITC
|
| TCL1
|
eBio1-21
|
6699
|
ICFC, WB, IHC
|
Purified, PE
|
| ZAP-70
|
1E7.2
|
6695
|
ICFC, WB, IP
|
Purified, Biotin, FITC, PE, PE-Cy5.5
|
FC = Fluorescence staining followed by flow cytometric analysis ICFC = Intracellular FC
IHC = Immunohistochemical staining IHC/F = IHC of frozen tissue IHC/P = IHC of paraffin embedded tissue
BA = Functional bioassays IP = Immunoprecipitation WB = Immunoblotting |
Table 2: Mouse Cancer Markers
|
Mouse Cancer Markers
|
| Mouse |
Clone |
SKU |
Applications |
Formats |
| B7-H1
|
MIH5
|
5982
|
FC, BA, IHC/F
|
Purified, FG Purified, Biotin, PE
|
|
|
1-111A
|
9971
|
FC
|
Purified, Biotin
|
| Bcl-xL
|
2H12
|
6994
|
WB
|
Purified
|
| DEC-205
|
205yekta
|
2051
|
FC, WB
|
Purified, Biotin, PE-Cy7
|
| E-Cadherin
|
DECMA-1
|
3249
|
FC, ELISA, IP, FA, IHC, WB
|
Purified, Alexa Fluor® 647
|
| EpCAM
|
G8.8
|
5791
|
FC, IH/F, IP
|
Purified, PE
|
| SLP-76
|
MS76
|
7761
|
ICFC, WB
|
Purified, FITC
|
| ZAP-70
|
1E7.2
|
6695
|
ICFC, WB, IP
|
Purified, Biotin, FITC, PE, PE-Cy5.5
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FC = Fluorescence staining followed by flow cytometric analysis ICFC = Intracellular FC
IHC = Immunohistochemical staining IHC/F = IHC of frozen tissue IHC/P = IHC of paraffin embedded tissue
BA = Functional bioassays IP = Immunoprecipitation WB = Immunoblotting |
Table 3: IL-12 Family
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IL-12 Family
|
| Specificity |
Description |
Clone |
SKU |
Formats |
| Human
|
IL-12 p40
|
C8.3
|
7127
|
Purified
|
|
|
IL-12 p35/p70
|
B-T21
|
7128
|
Purified
|
|
|
IL-12 p40/p70
|
C8.6
|
7129
|
Purified, FG Purified, Biotin, PE, Alexa Fluor® 488,
Alexa Fluor® 647, Pacific Blue®
|
|
|
IL-12 p70
|
-
|
8129
|
Recombinant, Carrier-free Recombinant
|
|
|
|
-
|
7126
|
ELISA Set
|
|
|
|
-
|
7929
|
ELISA Kit w/ pre-coated plates
|
|
|
|
-
|
7879
|
ELISPOT Set
|
|
|
IL-23
|
22A12
|
7238
|
Purified, FG Purified
|
|
|
|
-
|
8239
|
Recombinant, Carrier-free Recombinant
|
|
|
|
-
|
|
ELISA Set, ELISA Kit w/ pre-coated plates (please inquire)
|
|
|
IL-27
|
-
|
|
Recombinant (please inquire)
|
| Mouse
|
IL-12/IL-23 p40
|
C15.6
|
7125
|
Purified
|
|
|
|
C17.8
|
7123
|
FG Purified, Biotin, PE, Alexa Fluor® 647
|
|
|
IL-12 p35/p70
|
C18.2
|
7122
|
Purified
|
|
|
IL-12 p70
|
-
|
8121
|
Recombinant, Carrier-free Recombinant
|
|
|
|
-
|
7121
|
ELISA Set
|
|
|
|
-
|
7921
|
ELISA Kit w/ pre-coated plates
|
|
|
IL-12/IL-23 p40 (total)
|
-
|
7120
|
ELISA Set
|
|
|
|
-
|
7922
|
ELISA Kit w/ pre-coated plates
|
|
|
IL-23
|
G23-8
|
7232
|
Purified, FG Purified
|
|
|
|
-
|
8231
|
Recombinant, Carrier-free Recombinant
|
|
|
|
-
|
7234
|
ELISA Set
|
|
|
|
-
|
7231
|
ELISA Kit w/ pre-coated plates
|
|
|
IL-27 (EBI3 subunit)
|
please inquire
|
1272
|
Purified, Biotin
|
|
|
IL-27/IL-30 (p28)
|
please inquire
|
1301
|
Purified
|
|
|
IL-27
|
-
|
|
Recombinant (please inquire)
|
|
|
IL-27 (p28/EBI3)
|
-
|
7274
|
ELISA Set
|
| *Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. |
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