CD247 Molecule (CD247) antibody (Biotin)

Antigen: CD247 Molecule (CD247)

Synonyms: T3Z, CD3H, CD3Q, CD3Z, TCRZ, CD3-ZETA, Cd3, T3z, Cd3h, Cd3z, TCRk, Tcrz, CD3-eta, CD3zeta, AW552088, CD3-zeta, 4930549J05Rik, A430104F18Rik, CD247, im:7156386, LOC100136514, LOC100136549, LOC100305023

Clonality: Monoclonal (H57-597)

Host: Hamster

Reactivity: Mouse (Murine)

Conjugate: Biotin

Application: Flow Cytometry (FACS)

Catalog no.: ABIN114324

Additional Information

Alternative name: T Cell Receptor (TCR) alpha/beta

Immunogen: Affinity Purified D0-11. 10 TCRDonor: Armenian HamsterFusion Partner: Murine myeloma variant P3X63Ag. 653

Cross-Reactivity: Mouse (Murine)

Isotype: IgG

Clone: H57-597

Characteristics: Synonyms: TCRA, TCRB, T-Cell Receptor alpha, T-Cell Receptor beta, T-Cell Receptor alpha beta

Specificity: This mAb reacts with the surface of all ab TCR bearing cells and does not react withreceptors on gd TCR positive T cells. When used in an immobilized form, this antibody wasable to activate all ab TCR bearing T cell hybridomas tested to produce IL-2. Use of thisantibody in conjunction with an anti-CD3e mAb allows for accurate measurements of themutually exclusive sub-populations of ab TCR and gd TCR bearing T cells. Species: Mouse. Others not tested.

Application Details

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M, cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 1. 0 µg* of this Ab per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (Streptavidin-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/cCell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 1. 0 µg/10e6 cellsIsotypic Control: Biotin Hamster IgGCell Source Percentage of cells stained above control: Thymus: 62. 6% see FIGURE 1Splenic T Cells: 76. 2%Strain Distribution by Flow Cytometry Analysis: Cell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 1. 0 µg/106 cellsStrains Tested: BALB/c, C57BL/6, CBA/J, C3H/He, AKRPositive: BALB/c, C57BL/6, CBA/J, C3H/He, AKRNegative: none

Application Notes: Flow cytometry. Immunoprecipitation. This clone has also been reported to work with frozen sections and Western Blotting. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Concentration: 0.1 mg/ml

Buffer: PBS,0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring totalprotein concentration to 4-5 mg/ml.Label: Biotin

Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Research Area: Immunology, Receptors, Signaling

Restrictions: For Research Use only

Images

Figure 1

Publications

Palathumpat, Holm, Dejbakhsh-Jones et al.: "Treatment of BCL1 leukemia by transplantation of low density fractions of allogeneic bone marrow and spleen cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 148, Issue 10, pp. 3319-26, 1992 (PubMed).

Kubo, Born, Kappler et al.: "Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 142, Issue 8, pp. 2736-42, 1989 (PubMed).

Goodman, Lefrancois: "Intraepithelial lymphocytes. Anatomical site, not T cell receptor form, dictates phenotype and function." in: The Journal of experimental medicine, Vol. 170, Issue 5, pp. 1569-81, 1989 (PubMed).

Skarstein, Holmdahl, Johannessen et al.: "Oligoclonality of T cells in salivary glands of autoimmune MRL/lpr mice." in: Immunology, Vol. 81, Issue 4, pp. 497-501, 1994 (PubMed).

Paliwal, Ptak, Sperl et al.: "Recombinant soluble alphabeta T cell receptors protect T cells from immune suppression: requirement for aggregated multimeric, disulfide-linked alphabeta heterodimers." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 159, Issue 4, pp. 1718-27, 1997 (PubMed).