Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]

11 product images

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  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Lanes 1 - 6: Merged signal (red and green). Green - ab211017 observed at 40 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
  

  ab211017 was shown to react with Indoleamine 2, 3-dioxygenase in treated wild-type A549 cells in Western blot with no signal observed in treated IDO1 knockout cell line Human IDO1 (Indoleamine 2, 3-dioxygenase) knockout A549 cell line ab266949 (IDO1 knockout cell lysate ab256948). Wild-type A549 and IDO1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab211017 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution

  Lane 1: Wild-type A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate at 20 µg

  Lane 2: Wild-type A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate at 20 µg

  Lane 3: IDO1 knockout A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate at 20 µg

  Lane 4: IDO1 knockout A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate at 20 µg

  Lane 5: SK-OV-3 cell lysate at 20 µg

  Lane 6: MCF7 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 45 kDa

  Observed band size: 40 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
  

  Cytoplasmic and nuclear staining on dendritic cells of human spleen is observed (PMID: 21328335, PMID: 25271151).

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

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  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Exposure time: Lane 1-3: 3 minutes; Lane 4: 15 seconds.

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution

  Lane 1: Human ovary cancer lysate at 20 µg

  Lane 2: Human placenta lysate at 20 µg

  Lane 3: Human tonsil lysate at 20 µg

  Lane 4: SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 45 kDa

  Observed band size: 45 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
  

  Cytoplasmic and nuclear staining on dendritic cells of human tonsil is observed (PMID: 21328335, PMID: 25271151).

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

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  Immunocytochemistry/ Immunofluorescence - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-γ for 16 hours or untreated, labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
  

  The signal increased after treatment with IFN-γ (50 ng/ml) for 16 hours on HeLa cells.

  The nuclear counterstain is DAPI (blue).

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

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
  

  Cytoplasmic and nuclear staining on human endometrium cancer is observed (PMID: 26155395).

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

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  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

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

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution

  Lane 1: Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

  Lane 2: HeLa whole cell lysate treated with 50ng/ml Interferon-gamma (IFN-gamma, Recombinant Human Interferon gamma protein ab51240) for 16 hours at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 45 kDa

  Observed band size: 45 kDa

  Exposure time: 15s

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  Immunoprecipitation - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Indoleamine 2, 3-dioxygenase was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 50ng/ml IFN-γ for 16h with ab211017 at 1/40 dilution.
  

  Western blot was performed from the immunoprecipitate using ab211017 at 1/1000 dilution.

  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

  Lane 1: HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate 10 μg (Input).

  Lane 2: ab211017 IP in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab211017 in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 1 second.

  All lanes: Immunoprecipitation - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Predicted band size: 45 kDa

  Observed band size: 45 kDa

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  Flow Cytometry (Intracellular) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-? for 16h (red) or untreated (green), labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabeled 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.
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
  

  Cytoplasmic and nuclear staining on endothelial cells of human placenta is observed (PMID: 21328335, PMID: 25271151).

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)

  Tissue Microarrays stained for "Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]" using "ab211017"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab211017 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).