Staining of C57Bl/6 splenocytes with Anti-Human/Mouse CD45R (B220) APC (cat. 17-0452) and 0.06 μg of Anti-Mouse CD3e PerCP-eFluor® 710. Total viable cells were used for analysis.
Contents: Anti-Mouse CD3e PerCP-eFluor® 710 Catalog Number: 46-0033 Concentration: 0.2 mg/mL Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. Do not freeze. Light-sensitive material. Clone: eBio500A2 (500A2) Host/Isotype: Golden Syrian Hamster IgG
Product Options
Cat. No.
Size
Price
Add Qty to Cart
46-0033-80
25 ug
46-0033-82
100 ug
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
For research use only, not for diagnostic or therapeutic use.
Applications Tested
This eBio500A2 (500A2) antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.125 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.
PerCP-eFluor® 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative.
Our testing indicates that PerCP-eFluor® 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols.
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)
