Ly 2 microbeads

Manufactured by Miltenyi Biotec

Ly-2 microbeads are a type of magnetic particles used for cell separation techniques in research and clinical applications. They are made of a magnetic core surrounded by a polymer shell. The core function of Ly-2 microbeads is to enable the isolation and enrichment of specific cell populations from complex samples through magnetic separation.

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Lab products found in correlation

Complete rpmi, by Corning (1 mentions) Ova323 339, by GenScript (1 mentions) Macs ls column, by Miltenyi Biotec (1 mentions) Monocyte isolation kit, by Miltenyi Biotec (1 mentions) Ctla4 ig, by BioXCell (1 mentions) Pam3cys4, by InvivoGen (1 mentions)

Close spelling variants of this product

CD8a (Ly-2) MicroBeads (35 citations) Mouse CD8a (Ly-2) MicroBeads (8 citations) Anti-CD8 (Ly-2) microbeads (5 citations) CD8α (Ly-2) MicroBeads (5 citations) Anti‐CD8a (Ly‐2) MicroBeads (2 citations) Anti-CD8α (Ly-2) MicroBeads (2 citations) Anti-mouse-CD8-α (Ly-2) MicroBeads (2 citations)

5 protocols using ly 2 microbeads

Adoptive Transfer of Antigen-Specific CD8+ T Cells

Cited 5 times

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Donor mice were immunized with either PLP_178-191 or control OVA_323-339 (ISQAVHAAHAEINEAGR, GenScript, Piscataway, NJ) in CFA. At 15 days post-immunization, splenocytes and inguinal lymphocytes were harvested and as published previously (5 (link), 23 (link)–29 (link)), single cell suspensions were restimulated in vitro with 20μg/mL cognate antigen and 10pg/mL rIL-2 for 72h in complete RPMI (Corning, Tewksbury, MA). CD8 T cells were sorted to above 90% purity using Ly-2 microbeads (Miltenyi Biotech, Auburn, CA), and 5x10⁶ cells were adoptively transferred i.v. into recipient mice at times indicated. For mixed adoptive transfer experiments, CD8 T cells from cultures including equal inputs of bulk day 15-immune IFNγR−/− and IFNγ−/− cells (inguinal lymphocytes and splenocytes) were Ly-2 bead-sorted and transferred at 5x10⁶ per recipient. Unless otherwise specified, “control” groups consist of recipient mice injected i.v. with either PBS alone or OVA-CD8 donor cells, both of which have no effect on disease, as shown in this report and published previously (5 (link), 27 (link), 28 (link)). A 2:1 donor to recipient ratio was used in these experiments.

Boyden A.W., Brate A.A., Stephens L.M, & Karandikar N.J. (2020). Immune autoregulatory CD8 T cells require IFNγ responsiveness to optimally suppress CNS autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 205(2), 359-368.

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Adoptive Transfer of CD8 T Cells Mitigates EAE

Cited 1 time

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Mice were injected i.v. with 10⁷ cfu of recombinant LM. Seven days post-infection, these mice were immunized s.c. with emulsion containing Complete Freud’s Adjuvant supplemented with 200 μg Mycobacterium tuberculosis H37Ra (Difco, Detroit, MI) and 50–100 μg PLP_178–191 (ChinaPeptides) distributed over two sites on the flank. In all models, 250 ng pertussis toxin was administered i.p. on days 0 and 2 post-immunization. For CD8 T cell transfer, splenocytes were collected on day seven post- LM infection and reactivated with 20 μg/ml cognate antigen and 10 pg/ml recombinant human IL-2 (University of Iowa Hospitals and Clinics) for 72 h at 37 °C. CD8 T cells were magnetically isolated using Ly-2 microbeads (Miltenyi) and 10⁷ live cells were transferred to recipient mice i.v. After 24 h, active EAE was induced using PLP178-191/CFA immunization as described above. Clinical signs of EAE were assessed daily, and animals were scored based on previously defined criteria^{10 (link)}.

Itani F.R., Sinha S., Brate A.A., Pewe L.L., Gibson-Corley K.N., Harty J.T, & Karandikar N.J. (2017). Suppression of autoimmune demyelinating disease by preferential stimulation of CNS-specific CD8 T cells using Listeria-encoded neuroantigen. Scientific Reports, 7, 1519.

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Adoptive Transfer of Antigen-Specific CD8 T Cells

Cited 3 times

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Naïve mice were immunized with 50μg of PLP peptide (either P178 or P139) or OVA_323–339 in CFA. At day 14, spleens and inguinal lymph nodes were collected and reactivated in culture with cognate antigens and hIL-2 for 72hr as previously described (4 (link), 7 (link)–11 (link), 18 ). At the end of culture, cells were collected, and CD8 T cells were magnetically isolated using Ly-2 microbeads (Miltenyi) and 10×10⁶ live cells were transferred to recipient mice. Mice received CD8 T cell transfers i.v. prior to disease induction (d-1) or during the acute phase of disease (d11).

Brate A.A., Boyden A.W., Jensen I.J., Badovinac V.P, & Karandikar N.J. (2021). A functionally distinct CXCR3+/IFNγ+/IL-10+ subset defines disease-suppressive CNS-specific CD8 T cells. Journal of immunology (Baltimore, Md. : 1950), 206(6), 1151-1160.

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Adoptive Transfer of Activated OT-I T Cells

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Three days prior to adoptive transfer, spleen and inguinal lymph node from OT-I Tg mouse were crushed and cell suspensions were cultured in T-cell medium (45% RPMI-1640, 45% DMEM, 10% FBS, 1% Penicillin-Streptomycin, 1X 2-mercaptoethanol) with 0.5 μg/mL SIINFEKL and 10 ng/mL IL-2. After 2 days of culture, cell suspension was supplemented with fresh medium, SIINFEKL and IL-2. On day of transfer, CD8⁺ T cells were enriched from cell suspension using MACS LS column and Ly-2 microbeads (Miltenyi) and suspended in ice-cold sterile 1X PBS. Enrichment was verified (every 5^th experiment) to be >90%. 5 million activated T cells were transferred into mice by retro-orbital venous injection (i.v.) on days noted. T cell transfer was supplemented with five rIL-2 injections (20,000 IU per mouse, i.p.) every other day for 10 days.

Hegde S., Krisnawan V.E., Herzog B.H., Zuo C., Breden M.A., Knolhoff B.L., Hogg G.D., Tang J.P., Baer J.M., Mpoy C., Lee K.B., Alexander K.A., Rogers B.E., Murphy K.M., Hawkins W.G., Fields R.C., DeSelm C.J., Schwarz J.K, & DeNardo D.G. (2020). Dendritic cell paucity leads to dysfunctional immune surveillance in pancreatic cancer. Cancer cell, 37(3), 289-307.e9.

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Lung Transplant Immunomodulation Protocols

Cited 2 times

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Left orthotopic vascularized lung transplants were performed as previously described (10 (link)). Mice were treated with co-stimulatory blockade consisting of MR1 (250 μg intraperitoneally (i.p.)) and CTLA4-Ig (200 μg i.p.), on days 0 and 2, respectively (Bio X Cell, West Lebanon, NH). Pam₃Cys₄ was dissolved in distilled water at a concentration of 1 μg/μl and 50 μL were administered into the donor airway just prior to completing the bronchial anastomosis (Invivo Gen, San Diego, CA). In select experiments, LPS (20 μg) was dissolved in 50 μl of phosphate buffered saline and injected into the donor airway before connecting donor and recipient bronchi (Sigma, St. Louis, MO). The concentrations and administration route were based on previous studies examining the impact of Pam₃Cys₄ and LPS on pulmonary immune responses (12 (link)) (13 (link)) (14 (link)). For select experiments, 10x10⁶ CD8⁺ T cells (Ly-2 microbeads, Miltenyi Biotec, Bergisch Gladbach, Germany) (15 (link)) or 3x10⁶ monocytes (Monocyte Isolation Kit, Miltenyi Biotec) (16 (link)), isolated from the spleen or bone marrow, respectively, of naïve B6 WT mice were injected intravenously into B6 TLR2KO recipients at the time of transplantation.

Tanaka S., Gauthier J.M., Terada Y., Takahashi T., Li W., Hashimoto K., Higashikubo R., Hachem R.R., Bharat A., Ritter J.H., Nava R.G., Puri V., Krupnick A.S., Gelman A.E, & Kreisel D. (2020). Bacterial products in donor airways prevent the induction of lung transplant tolerance. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(1), 353-361.

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