47-0036-42

antibody from Invitrogen Antibodies

Targeting: CD247 CD3H, CD3Q, CD3Z

Provider product page for 47-0036-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
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Validations
Flow cytometry [1]
Other assay [4]

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Product number: 47-0036-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD3 Monoclonal Antibody (SK7), APC-eFluor™ 780, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The SK7 monoclonal antibody reacts with human and chimpanzee CD3e, a 20 kDa subunit of the TCR complex. Along with the other CD3 subunits gamma and delta, the epsilon chain is required for proper assembly, trafficking and surface expression of the TCR complex. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells. The SK7 and UCHT1 monoclonal antibodies cross-block binding, suggesting recognition of overlapping epitope. In contrast, clones OKT3 and SK7 see different epitopes. The antibody SK7 recognizes chimpanzee CD3. Applications Reported: This SK7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This SK7 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. APC-eFluor 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: SK7
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

CD247 protein structure - 47-0036-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Circulating percentages of Th17 cells and CCR6-positive cells in peripheral blood are increased in IVD degenerated patients when compared with controls. Heparinized peripheral whole blood cells from 20 patients and 15 healthy controls were stimulated with phorbol myristate acetate (PMA), ionomycin, and monensin for 4 h and subsequently stained with fluorochrome-labeled antibodies as described in Materials and Methods. A(a) Lymphocytes were gated by flow cytometry. A(b) CD3 + T subsets were gated by flow cytometry; the plots in the inset box represent the CD3 + T cells. A(c) Representative IL-17 expression levels in the CD3 + CD8 - T subsets (CD4 + T subsets) from each group are shown. The percentages of positive cells are shown in the upper left panels. A(d) Representative surface CCR6 expression levels on the CD3 + CD8 - IL-17 + subsets from each group are shown. The percentages of positive cells are shown in the right panel. (B) The percentage of circulating Th17 cells was significantly higher in IVD degenerated patients (2.973+-0.689%) than in the control group (1.039+-0.156%; *, p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 In vitro T cell proliferation. Mononuclear cells from psoriatic patients (red curve) and healthy controls (blue curve) were labelled with 1 uM CFSE prior to culturing and incubated for 5 days alone, with 1 ug/ml PHA ( A ) or in the presence of recombinant proteins K17 ( B ), S1 ( C ), S4 ( D ), PS1 ( E ) and PK ( F ) at concentration of 10 ug/ml. After 5 days, cells were labelled with a PE-conjugated anti-CD3 antibody and 7-AAD prior to flow cytometry analysis. Gated CD3+ lymphocytes are shown on the CFSE fluorescence histograms to demonstrate the decrease in fluorescence intensity during divisions. The higher peak for both patients and healthy controls represents a larger population of non-dividing parental cells (P). The signal of interest is the smaller peak that corresponds to dividing cells (shown using arrow under F ), and such cells are only observed in patents when treated with K17 ( B ), S1 ( C ), or S4 ( D ) proteins. Figures represent the percentage of CFSE low CD3+ cells (proliferating population).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2 Expression of PD-L1 on human HCC PLC/PRF/5 cells. (A) Human HCC PLC/PRF/5 cells were cultured alone in the absence of GPC3-CAR T cells in RPMI 1640 medium containing 10% FBS. (B) Human HCC PLC/PRF/5 cells were cocultured with GPC3-CAR T cells at an effector:Target ratio of 1:1 for 18 h in RPMI 1640 medium containing 10% FBS. PD-L1 was determined by flow cytometry in the CD3-negative gate, and the fixable, viable stain 780 was used for discriminating live from dead cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 4 Efficient disruption of PD-1 expression on the surface of GPC3-CAR T cells. PD-1 and CAR expression on the surface of T cells were detected by flow cytometry on day 3 after the re-stimulation with anti-CD3/anti-CD28 beads. UTD, untransduced T cells; WT, wild type.