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86603
CD3 (17A2) Rat mAb (FITC Conjugate)
Antibody Conjugates

CD3 (17A2) Rat mAb (FITC Conjugate) #86603

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
M Endogenous Rat IgG2b, kappa
Flow Cytometry

Flow cytometric analysis of live mouse splenocytes using CD3 (17A2) Rat mAb (FITC Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (FITC Conjugate) #56722 (dashed line).

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

For optimal flow cytometry results, we recommend 0.5 μg of antibody per test. A slight precipitate may be present, but will not interfere with the antibody performance. If precipitates are present, centrifuge the tube at 6,000xg for 10-30 sec. Draw off the supernatant and place into a light protective vial.

Application Dilutions
Flow Cytometry 1:100
Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

CD3 (17A2) Rat mAb (FITC Conjugate) recognizes endogenous levels of total CD3ε, CD3γ, and CD3δ proteins. This antibody detects epitopes within the extracellular domains.

Species Reactivity:

Mouse

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

This Cell Signaling Technology antibody is conjugated to FITC and tested in-house for direct flow cytometric analysis in mouse cells.

When T cells encounter antigens via the T cell receptor (TCR), information about the quantity and quality of antigens is relayed to the intracellular signal transduction machinery (1). This activation process depends mainly on CD3 (Cluster of Differentiation 3), a multiunit protein complex that directly associates with the TCR. CD3 is composed of four polypeptides: ζ, γ, ε and δ. Each of these polypeptides contains at least one immunoreceptor tyrosine-based activation motif (ITAM) (2). Engagement of TCR complex with foreign antigens induces tyrosine phosphorylation in the ITAM motifs and phosphorylated ITAMs function as docking sites for signaling molecules such as ZAP-70 and p85 subunit of PI-3 kinase (3,4). TCR ligation also induces a conformational change in CD3ε, such that a proline region is exposed and then associates with the adaptor protein Nck (5).

  1. Kuhns, M.S. et al. (2006) Immunity 24, 133-139.
  2. Pitcher, L.A. and van Oers, N.S. (2003) Trends Immunol. 24, 554-560.
  3. Osman, N. et al. (1996) Eur. J. Immunol. 26, 1063-1068.
  4. Hatada, M.H. et al. (1995) Nature 377, 32-38.
  5. Gil, D. et al. (2002) Cell 109, 901-912.
Entrez-Gene Id
12501 , 12502
Swiss-Prot Acc.
P22646 , P11942
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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Product # Size Price
86603S
100 µg $ 89.0