Anti-Granzyme A antibody [EPR29034-79]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Upper Left) / 293T (human embryonic kidney epithelial cell) transfected with an Granzyme A expression vector containing a myc-His-tag® (Upper Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Upper Right) / 293T cells transfected with homologous human Granzyme expression vector (GrzmB, GrzmH, GrzmK, GrzmM) containing a myc-His-tag® (Lower). cells labelling Granzyme A with ab321992 at 1/50 dilution (1ug) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Cells were first stained with rabbit IgG or ab321992. After fixation and permeability, cells stained with anti-myc tag conjugated to Alexa Fluor® 647.
Crossreactivity with protein Granzyme B, Granzyme H, Granzyme K, Granzyme M were fully tested.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with ab321992 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 dilution (Green).

Confocal image showing cytoplasmic staining in a subset of human PBMC (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ab195889 200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ab150081 1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD14 conjugated to Pacific blue.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) cells labelling Granzyme A with ab321992 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 dilution (Green).

Confocal image showing cytoplasmic staining in No-GFP-CD16.NK-92 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : 293T
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ab195889 200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ab150081 1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) (Right) / 293T (human embryonic kidney epithelial cell) (Left) cells labelling Granzyme A with ab321992 at 1/500 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Negative control : 293T.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD56 conjugated to PE.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Granzyme A antibody [EPR29034-79] (AB321992)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD8a conjugated to Pacific blue.