Clone EPR20129-217 (ab219803) has been successfully conjugated by Abcam. This image was generated using Anti-Granzyme B antibody [EPR20129-217] (PE). Please refer to ab225471 for protocol details.

Overlay histogram showing NK-92 cells stained with ab225471 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab225471, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20 mW blue solid-state laser (488nm) and 585/42 bandpass filter.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Granzyme B with ab208586 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on some stromal cells of human colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208586).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NK-92 (Human peripheral blood malignant Non-Hodgkin's lymphoma cell line) cells labeling Granzyme B with ab208586 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NK-92 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208586).

Flow cytometric analysis of 4% paraformaldehyde-fixed NK-92 (Human peripheral blood malignant Non-Hodgkin's lymphoma cell line) cells labeling Granzyme B with ab208586 at 1/500 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control(ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208586).

This IHC data was generated using the same anti-Granzyme B antibody clone [EPR20129-217] in a different buffer formulation (cat# ab208586).

Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling Granzyme B with ab208586 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on neutrophils and stroma cells of human cervix cancer is observed [PMID: 14512315]. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Clone EPR20129-217 (ab219803) has been successfully conjugated by Abcam. This image was generated using Anti-Granzyme B antibody [EPR20129-217] (Alexa Fluor® 488). Please refer to ab225472 for protocol details.

ab225472 staining Granzyme B in NK-92 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab225472 at 1/50 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Clone EPR20129-217 (ab219803) has been successfully conjugated by Abcam. This image was generated using Anti-Granzyme B antibody [EPR20129-217] (Alexa Fluor® 647). Please refer to ab225473 for protocol details.

ab225473 staining Granzyme B in NK-92 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab225473 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).