Anti granzyme a percp cy5

Manufactured by BioLegend

Anti-granzyme A-PerCp/Cy5.5 is a fluorescently labeled antibody that binds to the granzyme A protein. Granzyme A is a serine protease found in the cytotoxic granules of natural killer cells and cytotoxic T cells, and is involved in inducing cell death. The PerCp/Cy5.5 fluorescent label allows for detection and quantification of cells expressing granzyme A.

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Lab products found in correlation

Anti cd8 fitc clone sk1, by BioLegend (2 mentions) Bv510 streptavidin, by BioLegend (2 mentions) Anti cd62l pe cy5 clone dreg 56, by BioLegend (2 mentions) Anti cd3 af700 clonehit3a, by BioLegend (2 mentions) Anti cd45ra bv605, by BioLegend (2 mentions) Bv650 streptavidin, by R&D Systems (2 mentions) Anti granzyme k pe, by BioLegend (2 mentions) Anti granzyme m efluor660, by Thermo Fisher Scientific (2 mentions) Anti granzymeb pacific blue, by BioLegend (2 mentions) Near ir dead cell stain, by Thermo Fisher Scientific (1 mentions) Anti ki67 fitc, by BD (1 mentions) Anti cd4 viogreen, by Miltenyi Biotec (1 mentions) Anti granzymeb alexafluor700, by BD (1 mentions) Anti cd69 fitc, by Thermo Fisher Scientific (1 mentions) Anti perforin pacific blue, by BioLegend (1 mentions) Anti perforin fitc, by BD (1 mentions) Anti granzyme b apc, by Thermo Fisher Scientific (1 mentions) Anti t bet pe, by Santa Cruz Biotechnology (1 mentions) Anti cd8β pe, by Beckman Coulter (1 mentions) Macsquant, by Miltenyi Biotec (1 mentions)

3 protocols using anti granzyme a percp cy5

Assessing CD8+ T Cell Cytolytic Potential

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Rested ex vivo CD8⁺ T cells were assessed for CMV and
HIV specificity using HLA-matched tetramers. Tetramers were made via
biotinylated monomer conjugation to BV510 streptavidin (BioLegend). Monomers
were obtained from Dr. Soren Buus (University of Copenhagen). The A02-NV9
tetramer was used to stain for CMV-specificity while pools of A02-SL9, B27-KK10,
B53-QW9, B53-YY9, B57-IW9(p24), B57-IW9(RT), B57-KF11, B57-QW9, and B57-TW10
tetramers were used to stain for HIV-specificity. Subsequent surface stains
included anti-CD62L-PE-Cy5 (clone DREG-56; BioLegend), anti-CD3-AF700 (clone
HIT3a; BioLegend), anti-CD8-FITC (clone SK1; BioLegend), anti-CD45RA-BV605
(clone HI100; BioLegend), and LIVE/DEAD Blue. Following permeabilization, the
cells were intracellular stained using anti-perforin PE/Cy7 (clone B-D48;
BioLegend), anti-granzyme A-PerCp/Cy5.5 (clone CB9; BioLegend), anti-granzyme
B-Pacific Blue (clone GB11; BioLegend), anti-granzyme H (biotinylated and
secondary stained with BV650 streptavidin) (polyclonal antibody, R&D),
anti-granzyme K-PE (clone GM26E7; BioLegend), and anti-granzyme M-eFluor660
(clone 4B2G4; eBioscience). The cells were analyzed by flow cytometry.
Naïve CD8⁺ T cells within the mixed population of
CD8⁺ T cells were used as internal gating controls for
perforin and each granzyme as these cells do not express these effector
molecules.

Clayton K.L., Collins D.R., Lengieza J., Ghebremichael M., Dotiwala F., Lieberman J, & Walker B.D. (2018). Resistance of HIV-infected macrophages to CD8 T lymphocyte-mediated killing drives immune activation. Nature immunology, 19(5), 475-486.

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Multiparametric Flow Cytometry of Immune Cells

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For immunofluorescence staining, dead cells were excluded with the Near-IR Dead-Cell stain (Invitrogen). Antibodies used were: anti-CD3 PE-Cy7 or APC, anti-CD8 PerCP-Cy5.5 or eFluor 450, anti-CD69 FITC (eBioscience); anti-CD161 PE or APC, anti-CD4 VioGreen (Miltenyi Biotec); anti-Vα7.2 PE or FITC or PE-Cy7, anti-CD107α PE-Cy7, anti-Granzyme A PerCP-Cy5.5, anti-Perforin Pacific Blue, anti-granulysin PE, anti-FasL PE (BioLegend); anti-Granzyme B AlexaFluor700, anti-Perforin FITC, anti-Ki67 FITC (BD Biosciences), anti-Granzyme B APC (Invitrogen); anti-Granzyme K FITC (Immunotools); anti-T-bet PE (Santa Cruz Biotechnology); anti-Granzyme A FITC, anti-Blimp1 AlexaFluor488 (R&D); anti-CD8β PE (Beckman Coulter).
Data were collected on the flow cytometers LSRII (BD Biosciences) or MACSQuant (Miltenyi Biotec), and was analyzed using FlowJo v9.6 (TreeStar). For ImageStream analysis, see Supplementary Methods.

Kurioka A., Ussher J.E., Cosgrove C., Clough C., Fergusson J.R., Smith K., Kang Y.H., Walker L.J., Hansen T.H., Willberg C.B, & Klenerman P. (2014). MAIT cells are licensed through granzyme exchange to kill bacterially sensitized targets. Mucosal Immunology, 8(2), 429-440.

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Assessing CD8+ T Cell Cytolytic Potential

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