Product Information

Contents: Pacific Blue® anti-mouse CD3 (CD3e, epsilon subunit)
Catalog Number: 57-0033
Sizes: 25 ug, 100 ug
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃, BSA and/or gelatin for protein stabilization
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: eBio500A2 (500A2)
Isotype: Golden Syrian Hamster IgG

Staining of BALB/c splenocytes with APC anti-mouse CD19 (MB19-1) (cat. 17-0191) and staining buffer (autofluorescence) (left) or 0.5 μg of Pacific Blue® anti-mouse CD3e (500A2) (right). Cells in the lymphocyte gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-0033	Affinity Purified anti-mouse CD3 (CD3e, epsilon subunit)	N/A	N/A	FA FC IH/F
16-0033	Functional Grade* Purified anti-mouse CD3 (CD3e, epsilon subunit)	N/A	N/A	FA FC
13-0033	Biotin anti-mouse CD3 (CD3e, epsilon subunit)	N/A	N/A	FC
11-0033	Fluorescein isothiocyanate (FITC) anti-mouse CD3 (CD3e, epsilon subunit)	488	518	FC
12-0033	Phycoerythrin (PE) anti-mouse CD3 (CD3e, epsilon subunit)	488	575	FC
56-0033	Alexa Fluor® 700 anti-mouse CD3 (CD3e, epsilon subunit) (APC-Cy5.5 Equivalent)	633	723	FC
57-0033	Pacific Blue® anti-mouse CD3 (CD3e, epsilon subunit)	402	421	FC
*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 Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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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

For research use only, not for diagnostic or therapeutic use. This eBio500A2 (500A2) antibody has been reported for use in flow cytometric analysis.

The Pacific Blue® emits at 456 nm and can be excited with the Violet laser. Please make sure that your instrument is capable of detecting this fluorochome.

Applications Tested

This eBio500A2 (500A2) antibody has been tested by flow cytometric analysis. This can be used at less than or equal to 1 μg per million cells in a 100 μl total staining volume. 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)

Related Products

Cat. 57-4914 Pacific Blue® Golden Syrian Hamster IgG Isotype Control (clone n/a)

Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.