Staining of BALB/c splenocytes with 0.25 µg of Golden Syrian Hamster IgG Isotype Control Purified (cat. 14-4914) (open histogram) or 0.25 µg of Anti-Mouse CD3e Purified (filled histogram) followed by Anti-Syrian Hamster IgG FITC (cat. 11-4211). Cells in the lymphocyte gate were used for analysis.
Contents: Anti-Mouse CD3e Purified Catalog Number: 14-0033 Concentration: 0.5 mg/ml Formulation: aqueous buffer, 0.09% sodium azide, may contain carrier protein/stabilizer Storage Conditions: Store at 2-8°C. Clone: eBio500A2 (500A2) Host/Isotype: Golden Syrian Hamster IgG
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Cat. No.
Size
Price
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14-0033-81
50 ug
14-0033-82
100 ug
14-0033-85
500 ug
14-0033-86
1 mg
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The eBio500A2 monoclonal antibody reacts with the 25 kD epsilon subunit of the mouse CD3 complex. CD3 subunits γ, δ and ε are 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, as well as NKT cells. Binding of eBio500A2 to CD3 initiates the intracellular biochemical pathway resulting in cellular activation and proliferation. The 500A2 antibody is able to partially cross-block the 17A2 and 145-2C11 antibodies, indicating that all three of these anti-CD3 antibodies recognize distinct but overlapping epitopes.
Applications Reported
The eBio500A2 antibody has been reported for use in flow cytometric analysis. It has also been reported for use in in vitro activation of mouse T cells. (Please use Functional Grade purified eBio500A2 in functional assays).
Applications Tested
This eBio500A2 (500A2) antibody has been tested by flow cytometric analysis of mouse thymocyte and splenocyte suspensions. This can be used at less than or equal to 0.5 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 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Havran WL, Poenie M, Kimura J, Tsien R, Weiss A, Allison JP. Expression and function of the CD3-antigen receptor on murine CD4+8+ thymocytes. Nature. 1987 Nov 12-18;330(6144):170-3. (500A2, FC, FA, PubMed)
Portoles P, Rojo J, Golby A, Bonneville M, Gromkowski S, Greenbaum L, Janeway CA Jr, Murphy DB, Bottomly K. Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation. J Immunol. 1989 Jun 15;142(12):4169-75. (PubMed)
Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998 Feb 1;160(3):1158-65. (500A2, FC, PubMed)