Anti mouse cd8 antibody

Manufactured by BioLegend

Sourced in United States

The Anti-mouse CD8 antibody is a laboratory reagent used for the detection and analysis of CD8+ T cells in mouse samples. It specifically binds to the CD8 surface marker, which is expressed on a subset of T lymphocytes. This antibody can be used in various immunological techniques, such as flow cytometry, to identify and study CD8+ T cells.

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Alexa Fluor 594-conjugated anti-mouse CD8 antibody FITC-conjugated anti-mouse CD8 antibody PE anti-mouse CD8+ antibodies PE-Cy7-conjugated anti-mouse CD8 antibody PE)-conjugated anti-mouse CD8 antibody Phycoerythrin-CY7–conjugated anti-mouse CD8 antibody APC anti-mouse CD8 antibody Anti-mouse CD8 Anti-CD8 antibody CD8 antibody

3 protocols using anti mouse cd8 antibody

CD8+ T cells and NK depletion

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For CD8⁺ T cells depletion, 100 μg/mouse purified anti-mouse CD8 antibody (Biolegend, clone: 53-6.7, Cat# 100735) was injected via lateral tail vein 24 hours before the last vaccination. For NK depletion, 50 μg/mouse purified anti-Asialo-GM1 Antibody (Biolegend, clone: Poly21460, Cat# 146002) was injected through lateral tail vein 24 hours before vaccination.

Ren Y., Wang N., Hu W., Zhang X., Xu J, & Wan Y. (2015). Successive site translocating inoculation potentiates DNA/recombinant vaccinia vaccination. Scientific Reports, 5, 18099.

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Analyzing DC and T Cell Phenotypes

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As previously described [19 (link)], each group's DCs and T cells that were going to be examined in the coculture system were first collected, then washed twice with PBS after being digested with 0.25 percent trypsin. After this, the cells were tested. Following centrifugation, the supernatant was discarded, and the cells were resuspended in Eppendorf (EP) tubes at a concentration of 2 105 cells per 100 μL. Then, l μL of primary antibodies was added to the EP tube, including antimouse CD133 antibody (BioLegend, USA), antimouse CD11c antibody (BioLegend, USA), antimouse CD80 antibody (BioLegend, USA), antimouse CD86 antibody (BioLegend, USA), antimouse MHC-II antibody (BioLegend, USA), antimouse CD4 antibody (BioLegend, USA), antimouse CD8 antibody (BioLegend, USA), and antimouse FOXp antibody (BioLegend, USA). Following a thirty minute incubation period in the dark at a temperature of four degrees Celsius, the cells were resuspended in 0.2 milliliters of PBS before being washed twice with PBS. The phenotypic changes in DCs and T cells were identified with the help of flow cytometry (a BD FACSCalibur), and the analysis of the experiment's results was performed with the FlowJo software tool (FlowJo™ v10.7, http://www.flowjo.com).

Zhou H., Sun C., Li C., Hua S., Li F., Li R., Cai D., Zou Y., Cai Y, & Jiang X. (2022). The MicroRNA-106a/20b Strongly Enhances the Antitumour Immune Responses of Dendritic Cells Pulsed with Glioma Stem Cells by Targeting STAT3. Journal of Immunology Research, 2022, 9721028.

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Isolation and Transfer of ILC Precursors

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We pooled lungs from 10–12 (P1) newborn ZBTB16^CreGFP mice (on CD45.1 background). Single cells were stained with 7-AAD Viability staining solution (Biolegend), anti-mouse CD3ε antibody (145–2C11), anti–mouse CD4 antibody (GK1.5), anti-mouse CD8 antibody (53–5.8), anti–mouse CD11b antibody (M1/70), anti-mouse CD11c (N418), anti–mouse CD19 antibody (1D3), anti-mouse B220 (2FI), anti-mouse LY6G antibody (1A8), anti-mouse NKp46 antibody (29A1.4), anti-mouse α4β7 antibody (DATK32), anti-mouse CCR6 antibody (29–2L17), anti–mouse CD127 antibody (A7R34) and anti-mouse KLRG1 antibody (2FI) (all diluted 1:100, Biolegend). The stained cells were sorted with Sony SH800S. ZBTB16⁺ ILC precursors were sorted as Live CD45⁺Lineage (CD3ε, CD4, CD8, CD11b, CD11c, CD19, B220 and Ly6G), CD127⁺α4β7⁺NKp46⁻KLRG1⁻CCR6⁻CD25⁻PD1⁺GFP⁺ cells. Using this protocol, we isolated an enriched population of 800–1000 ZBTB16⁺ ILC precursors. We confirmed the purity of the sorted population by intracellular staining with anti-mouse RORγt antibody (Q31–378), anti-mouse T-bet antibody (4B10), anti-mouse PD1 antibody (J43), anti-mouse Id2 antibody (17-9475-82) and anti-mouse GATA3 antibody (16E10A23) (all diluted 1:50) (Supplementary Fig. 5 A, B). The cells were resuspended in normal saline to final concentration of 1×10³ cells/100 μl. We adoptively transferred 1×10³ ZBTB16⁺ ILC precursors via ntratracheal route.

Oherle K., Acker E., Bonfield M., Wang T., Gray J., Lang I., Bridges J., Lewkowich I., Xu Y., Ahlfeld S., Zacharias W., Alenghat T, & Deshmukh H. (2020). Insulin like growth factor 1 supports a pulmonary niche that promotes type 3 innate lymphoid cell development in newborn lungs.. Immunity, 52(2), 275-294.e9.

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