CD3 antibody (PerCP)

Catalog No. ABIN1112129

Product Details anti-CD3 Antibody (PerCP)

Target: CD3

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD3 antibody is conjugated to PerCP

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Characteristics: Monoclonal Mouse Anti-Human CD3 PerCP is recommended for use in flow cytometry for identification of T cells subsets in peripheral blood.

Clone: 33-2A3

Isotype: IgG2a

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells.

Comment:

Peridin-cholophyll-protein complex (Prozyme).

Sample Collection: 1. Transfer 100 µl of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (10^6 cells). 2. Add 20 µl of CD3 PerCP and mix gently with a vortex mixer. The 20 µl is a guideline only, the optimal volume should be determined by the individual laboratory. 3. The recommended negative control is a non-reactive PerCP-conjugated antibody of the same isotype. 4. Incubate in the dark at room temperature at 4°C for 30 minutes or at room temperature (20-25 °C) for 15 minutes. 5. Add 1,5 ml of Lysing Solution to each sample and mix gently with a vortex mixer. Incubate for 10 minutes at room temperature in the dark. 6. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 7. Add 2 ml 0.01 mol/l PBS (It betters that it containing 2% bovine serum albumin) and resuspend the cells by using a vortex mixer. 8. Centrifuge at 1000 x g for 5 minutes. Gently aspirate the supernatant and discard it leaving approximately 50 µl of fluid. 9. Resuspend pellet in an appropriate fluid for flow cytometry, e.g. 0.3 ml PBS. The PBS should contain 1% paraformaldehyde (fixative) if samples are not analysed the same day. 10. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis. Samples can be run up to 24 hours after lysis.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer containing 1% bovine serum albumin (BSA) and 0,09% Sodium azide, pH 7.2.

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Storage: 4 °C

References for anti-CD3 antibody (PerCP) (ABIN1112129)

Gimferrer, Calvo, Mittelbrunn, Farnós, Sarrias, Enrich, Vives, Sánchez-Madrid, Lozano: "Relevance of CD6-mediated interactions in T cell activation and proliferation." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 173, Issue 4, pp. 2262-70, (2004) (PubMed).

Gimferrer, Farnós, Calvo, Mittelbrunn, Enrich, Sánchez-Madrid, Vives, Lozano: "The accessory molecules CD5 and CD6 associate on the membrane of lymphoid T cells." in: The Journal of biological chemistry, Vol. 278, Issue 10, pp. 8564-71, (2003) (PubMed).

Bohnhorst, Bjørgan, Thoen, Natvig, Thompson: "Bm1-Bm5 classification of peripheral blood B cells reveals circulating germinal center founder cells in healthy individuals and disturbance in the B cell subpopulations in patients with primary Sjögren's syndrome." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 167, Issue 7, pp. 3610-8, (2001) (PubMed).

Target Details for CD3

Target: CD3

Alternative Name: CD3 (CD3 Products)

Background: This antibody recognizes 22/26/30 kDa MW lymphocyte surface molecules associated with the T cell antigen receptor complex. Reacts with 85% of peripheral blood T lymphocytes, 70% of thymocytes, the majority of T cell chronic lymphocytic leukaemias, Sezary leukaemias and approximately 70% of acute lymphoblastic leukaemias of T cell origin.

Pathways: TCR Signaling, Ubiquitin Proteasome Pathway