Anti-Maus CD45R/B220 Magnetic Particles

Produktnummer ABIN1305233

Produktdetails

Target: CD45 (PTPRC) (Protein tyrosine Phosphatase, Receptor Type, C (PTPRC))

Reaktivität: Maus

Wirt: Ratte

Klonalität: Monoklonal

Konjugat: Magnetic Particles

Applikation: Separation (Sep)

Marke: BD IMag™

Anwendungsinformationen

Protokoll: 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, 2mM EDTA, and 0.09% sodium azide. Place on ice. Although our experience indicates that 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 CD45R/B220 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

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

Konservierungsmittel: Sodium azide

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

Lagerung: 4 °C

Informationen zur Lagerung: Store undiluted at 4° C. Page 1 of 3551513 Rev. 1

Referenzen

Kobata, Takasaki, Asahara, Hong, Masuko-Hongo, Kato, Hirose, Shirai, Kayagaki, Yagita, Okumura, Nishioka: "Apoptosis with FasL+ cell infiltration in the periphery and thymus of corrected autoimmune mice." in: Immunology, Vol. 92, Issue 2, pp. 206-13, (1998) (PubMed).

Sagara, Sugaya, Tokoro, Tanaka, Takano, Kodama, Nakauchi, Takahama: "B220 expression by T lymphoid progenitor cells in mouse fetal liver." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 158, Issue 2, pp. 666-76, (1997) (PubMed).

Renno, Hahne, Tschopp, MacDonald: "Peripheral T cells undergoing superantigen-induced apoptosis in vivo express B220 and upregulate Fas and Fas ligand." in: The Journal of experimental medicine, Vol. 183, Issue 2, pp. 431-7, (1996) (PubMed).

Rolink, ten Boekel, Melchers, Fearon, Krop, Andersson: "A subpopulation of B220+ cells in murine bone marrow does not express CD19 and contains natural killer cell progenitors." in: The Journal of experimental medicine, Vol. 183, Issue 1, pp. 187-94, (1996) (PubMed).

Laouar, Ezine: "In vivo CD4+ lymph node T cells from lpr mice generate CD4-CD8-B220+TCR-beta low cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 153, Issue 9, pp. 3948-55, (1994) (PubMed).

Ballas, Rasmussen: "Lymphokine-activated killer cells. VII. IL-4 induces an NK1.1+CD8 alpha+beta- TCR-alpha beta B220+ lymphokine-activated killer subset." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 150, Issue 1, pp. 17-30, (1993) (PubMed).

Hathcock, Hirano, Murakami, Hodes: "CD45 expression by B cells. Expression of different CD45 isoforms by subpopulations of activated B cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 149, Issue 7, pp. 2286-94, (1992) (PubMed).

Hardy, Carmack, Shinton, Kemp, Hayakawa: "Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow." in: The Journal of experimental medicine, Vol. 173, Issue 5, pp. 1213-25, (1991) (PubMed).

Coffman: "Surface antigen expression and immunoglobulin gene rearrangement during mouse pre-B cell development." in: Immunological reviews, Vol. 69, pp. 5-23, (1983) (PubMed).

Antigendetails

Target: CD45 (PTPRC) (Protein tyrosine Phosphatase, Receptor Type, C (PTPRC))

Andere Bezeichnung: CD45R/B220 (PTPRC Produkte)

Synonyme: PTPRC Accessory Reagents, si:dkeyp-47c5.1 Accessory Reagents, B220 Accessory Reagents, CD45 Accessory Reagents, CD45R Accessory Reagents, GP180 Accessory Reagents, L-CA Accessory Reagents, LCA Accessory Reagents, LY5 Accessory Reagents, T200 Accessory Reagents, Cd45 Accessory Reagents, Ly-5 Accessory Reagents, Lyt-4 Accessory Reagents, loc Accessory Reagents, Lca Accessory Reagents, RT7 Accessory Reagents, cd45 Accessory Reagents, protein tyrosine phosphatase, receptor type C Accessory Reagents, protein tyrosine phosphatase, receptor type, C Accessory Reagents, receptor-type tyrosine-protein phosphatase C Accessory Reagents, PTPRC Accessory Reagents, ptprc Accessory Reagents, LOC100074271 Accessory Reagents, Ptprc Accessory Reagents

Hintergrund: BD IMag™ anti-mouse CD45R/B220 Particles - DM are magnetic nanoparticles that have monoclonal antibody conjugated to their surfaces. These particles are optimized for the positive selection or depletion of CD45R/B220-bearing leukocytes using the BD IMagnet™. CD45R/B220 is reportedly expressed on B lymphocytes at all stages from pro-B through mature and activated B cell. It is also has been reported to be found on abnormal T cells involved in the lymphadenopathy of lpr/lpr and gld/gld mutant mice, on lytically active subsets of lymphokine-activated killer cells (NK cells and non-MHC-restricted CTL), on apoptotic T lymphocytes of mice injected with bacterial superantigen, on a population of NK-cell precursors in the bone marrow, and on B-lymphocyte, T-lymphocyte, and macrophage progenitors in fetal liver. The CD45R/B220 antigen is reportedly not on hematopoietic stem cells, plasma cells, resting T lymphocytes, or MHC-restricted CTL. Leukocytes are labeled with BD IMag™ anti-mouse CD45R/B220 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.