Anti cd57

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-CD57 is a laboratory reagent used for the identification and enumeration of CD57-positive cells in flow cytometry analysis. It is a monoclonal antibody that binds to the CD57 antigen expressed on a subset of natural killer cells and a small population of T cells. The core function of Anti-CD57 is to serve as a tool for cellular immunophenotyping and research applications.

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Anti-CD57-FITC (8 citations) FITC-conjugated anti-human CD57 (3 citations)

3 protocols using anti cd57

Characterization of Senescent and Exhausted T Cells

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PBMCs were isolated using Ficoll-Paque (GE Healthcare, USA), and the cellular phenotypic of senescent and exhausted T cells was analyzed by flow cytometry. For surface markers analysis, cells were stained in PBS containing 2% fetal bovine serum (FBS, Thermo fisher, USA) with antibodies as indicated. Then, flow cytometric analysis was carried out in BD LSRFortessa X20. The gating strategy to identify T cell subsets was applied as described previously (27 (link)). Antibodies used in this study were purchased from BD Biosciences and eBioscience, including anti-CD4 (GK1.5, 1:100), anti-CD8 (53-6.7, 1:100), anti-CD25 (PC61, 1:100), anti-CD45RA (HI100, 1:100), anti-CXCR3(G025H7, 1:100), anti-CCR4 (L291H4, 1:100), anti-CCR6 (G034E3, 1:100), anti-CCR7 (G043H7, 1:100), anti-CD127 (A019D5, 1:100), anti-CXCR5 (RF8B2, 1:100), anti-CD28 (CD28.2, 1:100), anti-CD57 (NK-1, 1:100), anti-KLRG1 (2F1, 1:100), anti-PD-1 (EH12.2H7, 1:100), anti-TIM3 (F38-2E2, 1:100), and fixable viability dye eFluor 780 (eBioscience, Cat#65-0865-14, 1:1,000).

Tang X., Deng B., Zang A., He X., Zhou Y., Wang D., Li D., Dai X., Chen J., Zhang X., Liu Y., Xu Y., Chen J., Zheng W., Zhang L., Gao C., Yang H., Li B, & Wang X. (2022). Characterization of age-related immune features after autologous NK cell infusion: Protocol for an open-label and randomized controlled trial. Frontiers in Immunology, 13, 940577.

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Detailed Immune Cell Characterization

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Lymphocytes from human blood/tumor samples were incubated for 30 min with the following mouse anti-human monoclonal antibodies: anti-CD45, anti-CD11b and anti-NKp80 (Biolegend, San Diego, CA, USA); anti-CD3, anti-CD56, anti-CD27, anti-CD7, anti-CD2, anti-CD57, anti-CD11c, anti-CD117 and anti-CD127 (eBioscience, San Diego, CA, USA); anti-NKG2D, anti-NKp30, anti-CD226 and anti-NKp46 (BD Bioscience, San Jose, CA, USA); anti-NKG2A (R&D Systems Inc, Minneapolis, MN, USA); and KIR2DL1/DS1, KIR2DL2/DL3/DS2, and KIR3DL1/DS1 (Beckman Coullter, Fullerton, CA, USA). Mouse serum was used to block non-specific Fc receptor binding, and isotype-matched IgGs were used as negative control antibodies. The samples were analyzed using a fluorescence activating cell sorter Calibur flow cytometer (BD Biosciences). Flow cytometry data were analyzed using the FlowJo software (Tree Star, Inc., Ashland, OR, USA).

Zhang Q.F., Yin W.W., Xia Y., Yi Y.Y., He Q.F., Wang X., Ren H, & Zhang D.Z. (2016). Liver-infiltrating CD11b−CD27− NK subsets account for NK-cell dysfunction in patients with hepatocellular carcinoma and are associated with tumor progression. Cellular and Molecular Immunology, 14(10), 819-829.

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Phenotypic Analysis of Senescent and Exhausted T Cells

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Peripheral blood mononuclear cells (PBMCs) were isolated using coll-paque (GE Healthcare, USA) for cellular phenotypic analysis of senescent and exhausted T cells by ow cytometry. For analysis of surface markers, cells were stained in PBS containing 2% fetal bovine serum (FBS, Thermo sher, USA)
with antibodies as indicated. Then ow cytometric analysis was carried out in BD LSRFortessa X20. The gating strategy to identify T cell subsets was applied as described previously 28 . Brie y, markers related to T cell senescence and exhaustion were also monitored using PD-1 TIM3 CD28 and CD57 antibodies.
Antibodies used in this study were from BD Biosciences and eBioscience, including anti-CD4 (GK1.5, 1:100), anti-CD8 (53-6.7, 1:100), anti-CD25 (PC61, 1:100), anti-CD45RA (HI100, 1:100), anti-CXCR3 (G025H7, 1:100), anti-CCR4 (L291H4, 1:100), anti-CCR6 (G034E3, 1:100), anti-CCR7 (G043H7, 1:100), anti-CD127 (A019D5, 1:100), anti-CXCR5 (RF8B2, 1:100), anti-CD28 (CD28.2, 1:100), anti-CD57 (NK-1, 1:100), anti-KLRG1 (2F1, 1:100), anti-PD-1 (EH12.2H7, 1:100), anti-TIM3 (F38-2E2, 1:100).

Tang X., Deng B., Zang A., He X., Zhou Y., Wang D., Li D., Zhang X., Liu Y., Yong-hua X., Chen J., Zheng W., Zhang L., Gao C., Yang H., Li B., & Wang L. (2021). Characterization Of Age-Related Immune Features After Autologous NK Cell Infusion.

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