Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (AB281580)

Immunohistochemistry analysis of paraffin-embedded human cervix carcinoma tissue staining Indoleamine 2, 3-dioxygenase using ab281580 at a 1/50 diluiton (10.0 μg/ml).

Positive staining on human cervix carcinoma.

The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab281580 at 1/50 dilution (shown in Red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.
Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (AB281580)

Immunocytochemistry analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells treated with human Interferon-γ (50 ng/ml) for 16 hours, labeling Indoleamine 2, 3-dioxygenase with ab281580 at 1/50 (10.0 μg/ml) dilution (Red).

Confocal image showing cytoplasmic and nuclear staining in Interferon-γ treated HeLa cells.

ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 dilution (Green). The nuclear counterstain was DAPI (Blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (AB281580)

Flow Cytometry analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) treated with 50ng/ml Human Interferon-γ for 16 hours (Red)/ Untreated control (Green) cells stained for Indoleamine 2, 3-dioxygenase using ab281580 at a 1/500 dilution (0.1μg) (red), compared to Rabbit monoclonal isotype control - Alexa Fluor® 647 (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

The level of Indoleamine 2, 3-dioxygenase expression can be induced by IFN-γ treatment (PMID : 16368976).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (AB281580)

Immunohistochemistry analysis of paraffin-embedded human spleen tissue staining Indoleamine 2, 3-dioxygenase using ab281580 at a 1/50 diluiton (10.0 μg/ml).

Positive staining on human spleen.

The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab281580 at 1/50 dilution (shown in Red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.
Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).