Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (AB289888)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Granzyme B with ab289888 at 1/100 dilution, followed by ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Negative control : no staining on the glomerulus and tubule epithelia in human kidney (PMID : 28900605).

The section was incubated with ab289888 for 30 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (AB289888)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Granzyme B with ab289888 at 1/100 dilution, followed by ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human tonsil.

The section was incubated with ab289888 for 30 mins at room temperature and followed by rat IgG antibody (ab102248) for 8 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection).

• WB

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Western blot - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (AB289888)

Negative control : Jurkat (PMID : 18437383, 25168906

5% NFDM/TBST was used as a blocking and diluting buffer.

All lanes:

Western blot - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (ab289888) at 1/1000 dilution

Lane 1:

KARPAS-299 (human anaplastic large cell lymphoma), whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 3:

SR (human pleural effusion lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

false

Exposure time: 48s

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) (AB289888)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized No-GFP-CD16.NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells labelling Granzyme B with ab289888 at 1/50 dilution (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rat IgG Fc (DyLight 488, ab96971) at 1/2000 dilution was used as the secondary antibody.