Anti-Souris CD90.2 (Thy1.2) Magnetic Particles

N° du produit ABIN1305246

Détail du produit

Antigène: CD90.2 / Thy-1.2

Reactivité: Souris

Hôte: Rat

Clonalité: Monoclonal

Conjugué: Magnetic Particles

Application: Separation (Sep)

Marque: BD IMag™

Information d'application

Protocole: 1. Prepare a single-cell suspension from the lymphoid tissue of interest according to standard laboratory procedures. Remove clumps of cells and/or debris by passing the suspension through a 70-µm nylon cell strainer.
2. Dilute BD IMag™ Buffer (10X) (Cat. No. 552362) 1:10 with sterile distilled water or prepare 1X BD IMag™ buffer by supplementing Phosphate Buffered Saline with 0.5% BSA, 2 mM EDTA, and 0.09% sodium azide). Place on ice. Although our experience indicates that the use of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141) is not required for optimal cell separation, some laboratories may want to use it in their studies. If adding Mouse BD Fc Block™, proceed to Step 3. If not adding Mouse BD Fc Block™, proceed to Step 4.
3. Add Mouse BD Fc Block at 0.25 µg/10e6 cells, and incubate on ice for 15 minutes.
4. Wash cells with at least an equal volume of 1X BD IMag™ buffer, and carefully aspirate all the supernatant.
5. Vortex the BD IMag™ anti-mouse CD90.2 Particles - DM thoroughly, and add 50 µl of particles for every 10e7 total cells.
6. MIX THOROUGHLY. Refrigerate at 6°C - 12°C for 30 minutes.
7. Bring the BD IMag-particle labeling volume up to 1 - 8 x 10e7 cells/ml with 1X BD IMag™ buffer, and immediately place the tube on the BD IMagnet™. Incubate at room temperature for 6 - 8 minutes.
8. With the tube on the BD IMagnet™, carefully aspirate off the supernatant. This supernatant contains the negative fraction.
9. Remove the tube from the BD IMagnet™, and add 1X BD IMag™ buffer to the same volume as in Step 7. Gently resuspend cells by pipetting briefly, and return the tube to the BD IMagnet™ for another 2 - 4 minutes.
10. With the tube on the BD IMagnet™, carefully aspirate off the supernatant and discard.
11. Repeat Steps 9 and 10.
12. After the final wash step, resuspend the positive fraction in an appropriate buffer and at an appropriate concentration for further analysis.
NOTE: Avoid nonspecific labeling by working quickly and adhering to recommended incubation times.

Restrictions: For Research Use only

Stockage

Format: Liquid

Buffer: Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: 4 °C

Stockage commentaire: Store undiluted at 4° C. Page 1 of 3551518 Rev. 5

Référence

Zheng, Han, Kelsoe: "T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1." dans: The Journal of experimental medicine, Vol. 184, Issue 3, pp. 1083-91, (1997) (PubMed).

Radrizzani, Carminatti, Pivetta, Idoyaga Vargas: "Developmental regulation of Thy 1.2 rate of synthesis in the mouse cerebellum." dans: Journal of neuroscience research, Vol. 42, Issue 2, pp. 220-7, (1996) (PubMed).

Ikuta, Uchida, Friedman, Weissman: "Lymphocyte development from stem cells." dans: Annual review of immunology, Vol. 10, pp. 759-83, (1992) (PubMed).

Lefrançois: "Extrathymic differentiation of intraepithelial lymphocytes: generation of a separate and unequal T-cell repertoire?" dans: Immunology today, Vol. 12, Issue 12, pp. 436-8, (1992) (PubMed).

Phipps, Penney, Keng, Quill, Paxhia, Derdak, Felch: "Characterization of two major populations of lung fibroblasts: distinguishing morphology and discordant display of Thy 1 and class II MHC." dans: American journal of respiratory cell and molecular biology, Vol. 1, Issue 1, pp. 65-74, (1990) (PubMed).

Tigelaar, Lewis, Bergstresser: "TCR gamma/delta+ dendritic epidermal T cells as constituents of skin-associated lymphoid tissue." dans: The Journal of investigative dermatology, Vol. 94, Issue 6 Suppl, pp. 58S-63S, (1990) (PubMed).

Ledbetter, Herzenberg: "Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens." dans: Immunological reviews, Vol. 47, pp. 63-90, (1980) (PubMed).

Ledbetter, Rouse, Micklem, Herzenberg: "T cell subsets defined by expression of Lyt-1,2,3 and Thy-1 antigens. Two-parameter immunofluorescence and cytotoxicity analysis with monoclonal antibodies modifies current views." dans: The Journal of experimental medicine, Vol. 152, Issue 2, pp. 280-95, (1980) (PubMed).

Détail du antigène

Antigène: CD90.2 / Thy-1.2

Autre désignation: CD90.2 (Thy1.2) (CD90.2 / Thy-1.2 Produits)

Synonymes: CD90 Accessory Reagents, T25 Accessory Reagents, Thy-1 Accessory Reagents, Thy-1.2 Accessory Reagents, Thy1.1 Accessory Reagents, Thy1.2 Accessory Reagents, thymus cell antigen 1, theta Accessory Reagents, Thy1 Accessory Reagents

Sujet: BD IMag™ anti-mouse CD90.2 (Thy-1.2) Particles - DM are magnetic nanoparticles that have monoclonal antibody conjugated to their surfaces. These particles are optimized for the positive selection or depletion of CD90.2-bearing leukocytes using the BD IMagnet™. The CD90.2 alloantigen is expressed on thymocytes, most peripheral T lymphocytes, some intraepithelial T lymphocytes (IEL, DEC), epithelial cells, fibroblasts, neurons, hematopoietic stem cells, but not B lymphocytes, of most mouse strains. 30-H12 mAb has been reported not to cross-react with mouse Thy-1.1 (e.g., AKR/J, PL), or with rat Thy-1. Leukocytes are labeled with BD IMag™ anti-mouse CD90.2 Particles - DM according to the Magnetic Labeling Protocol. This labeled cell suspension is then placed within the magnetic field of the BD IMagnet™ (Cat. No. 552311). Labeled cells migrate toward the magnet (positive fraction), leaving the unlabeled cells in suspension so they can be drawn off (negative fraction). The tube is then removed from the magnetic field for resuspension of the positive fraction. The separation is repeated twice to increase the purity of the positive fraction. The magnetic separation steps are diagrammed in the Separation Flow Chart. After the positive fraction is washed, the small size of the magnetic particles allows the positive fraction to be further evaluated in downstream applications such as flow cytometry.

Synonyms: Thy-1.2