Cd8a ly 2

Manufactured by Miltenyi Biotec

Sourced in United States

CD8a (Ly-2) is a lab equipment product. It is a marker for the CD8+ T cell subset.

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Lab products found in correlation

Carboxyfluorescein succinimidyl ester (cfse), by GenScript (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Cd4 l3t4, by Miltenyi Biotec (1 mentions) Macs ld column, by Miltenyi Biotec (1 mentions) 4 6 diamidino 2 phenylindole dapi, by BD (1 mentions) Facscan, by BD (1 mentions) Siinfekl, by Merck Group (1 mentions) Siinfekl peptide, by Thermo Fisher Scientific (1 mentions) Incomplete freund s adjuvant, by Merck Group (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Quadromacs separator, by Miltenyi Biotec (1 mentions) Cd11c microbeads, by Miltenyi Biotec (1 mentions)

3 protocols using cd8a ly 2

In Vitro CD8+ T-Cell Cytotoxicity Assay

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Splenocytes (5 × 10⁷ cells/ml) from naive C57BL/6 mice were divided into two populations, labeled with two different concentrations of CFSE (Life Technology, USA). One population was pulsed with 4 μg/ml of NP_366–374 peptide (GenScript, Netherlands) for 1 h at 37°C and treated for 15 min at 37°C with 0.5 μM CFSE (CFSE^low). The other population remained un-pulsed and was treated with 5 μM CFSE (CFSE^high). The CFSE^low (NP loaded) and CFSE^high cells (control) were mixed at a 1:1 ratio, washed twice in PBS + 2% FBS and incubated for 16 h with CD8+ T-cells obtained from the spleen or lung of animals immunized with OVX836, and purified by positive selection using CD8a (Ly-2) (Miltenyi Biotec, USA). This assay allows for the measurement of the intrinsic capacity of CD8+ T-cells to kill target cells to determine the actual value of cell specific lysis: 1) Ratio = %[CFSE^high] peak/%[CFSE^low] peak, 2) Percent Specific Lysis = [1 − (Control ratio/Experimental ratio)] × 100, as described previously (20 (link)).

Del Campo J., Bouley J., Chevandier M., Rousset C., Haller M., Indalecio A., Guyon-Gellin D., Le Vert A., Hill F., Djebali S., Leverrier Y., Marvel J., Combadière B, & Nicolas F. (2021). OVX836 Heptameric Nucleoprotein Vaccine Generates Lung Tissue-Resident Memory CD8+ T-Cells for Cross-Protection Against Influenza. Frontiers in Immunology, 12, 678483.

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Comprehensive Analysis of Leukemic Cells

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Flow cytometry analysis was performed as described previously(17 (link)), using a FACScan (BD Pharmingen) instrument. The antibodies used to characterize leukemic cells are indicated in supplementary Table S2. 4′,6-diamidino-2-phenylindole (DAPI) (BD Pharmingen, Cat#564907) or propidium iodide (PI) (Invitrogen, Cat# P3566) were used for viability staining.
For sorting DN cells; DN thymocytes were enriched by staining with CD4 (L3T4)(Cat# 130–117-043) and CD8a (Ly-2)(Cat#130–117-044) Microbeads (Miltenyi biotec), and depletion of the stained cells using MACS LD column (Cat# 130–042-901, Miltenyi biotec), according to the manufacturer’s recommended protocol. The depleted cells were then stained with mouse anti CD4, CD8, CD25, CD44 specific antibodies and sorted with an Aria FACS Aria II, FACS instrument.
Tissues were fixed in 10% Formalin, paraffin-embedded, and sections stained with hematoxylin and eosin, CD3 (MCA1477, AbD Serotec, Raleigh, NC, USA), B220 (553086, BD Pharmingen), or Myeloperoxidase (1:1000, DAKO-A0398). Tissue histology was analyzed as described previously(18 (link)).

Goldberg L., Negi V., Chung Y.J., Onozawa M., Zhu Y.J., Walker R.L., Pierce R., Patel D.P., Krausz K.W., Gonzalez F.J., Goodell M.A., Rodriguez B.A., Meltzer P.S, & Aplan P.D. (2021). Mutant Idh2 cooperates with a NUP98-HOXD13 fusion to induce early immature thymocyte precursor ALL. Cancer research, 81(19), 5033-5046.

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Isolation and Activation of Immune Cells for In Vivo Studies

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Isolation of CD8⁺ T cells or CD11c⁺ DCs was performed by immunomagnetic selection from splenocytes using Manual MACS Cell Separation Technology (QuadroMACS Separator, LS columns, CD8a [Ly-2] and CD11c MicroBeads; Miltenyi Biotec), according to the manufacturer’s instructions. For in vitro experiments, SIINFEKL peptide (Invitrogen) was added at a concentration of 5 μM. In competitive in vivo experiments, 5 × 10⁵ to 1 × 10⁶ each of T^CXCR4 and T^Control were mixed into a 1:1 ratio before injection, and vaccination was performed at 24 hours and on day 29 by s.c. injection of 200 μM SIINFEKL or an irrelevant peptide in a 1:1 ratio with incomplete Freund’s adjuvant (Sigma-Aldrich). Alternatively, mice received 1 × 10⁶ CD11c⁺ peptide-loaded DCs intravenously.

Khan A.B., Carpenter B., Santos e Sousa P., Pospori C., Khorshed R., Griffin J., Velica P., Zech M., Ghorashian S., Forrest C., Thomas S., Gonzalez Anton S., Ahmadi M., Holler A., Flutter B., Ramirez-Ortiz Z., Means T.K., Bennett C.L., Stauss H., Morris E., Lo Celso C, & Chakraverty R. (2018). Redirection to the bone marrow improves T cell persistence and antitumor functions. The Journal of Clinical Investigation, 128(5), 2010-2024.

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