Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89]

14 product images

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  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Western blot: Anti-IDO1 antibody [EPR28349-89] (ab311847) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab311847 was shown to bind specifically to IDO1. A band was observed at 42 kDa in treated wild-type A549 cell lysates with no signal observed at this size in IDO1 knockout cell line. To generate this image, wild-type and IDO1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847) at 1/1000 dilution

  Lane 1: Wild-type A549 Treated IFN gamma (25 ng/mL, 48 h) ab281500 cell lysate at 20 µg

  Lane 2: Wild-type A549 Vehicle Control IFN gamma (0 ng/mL, 48 h) ab255554 cell lysate at 20 µg

  Lane 3: IDO1 knockout A549 Treated IFN gamma (25 ng/mL, 48 h) ab281489 cell lysate at 20 µg

  Lane 4: IDO1 knockout A549 Vehicle Control IFN gamma (0 ng/mL, 48 h) ab263515 cell lysate at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 45 kDa

  Observed band size: 42 kDa

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  Flow Cytometry (Intracellular) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype control (Left) /RAW 264.7 (mouse Abelson murine leukaemia virus-induced tumor macrophage) treated with 50ng/ml IFN-? for 24h (Middle) / Untreated RAW 264.7 (Right) cells labelling Indoleamine 2, 3-dioxygenase with ab311847 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with an IDO1 expression vector containing a his tag. No staining on (B) HEK-293T transfected with IDO2 expression vector containing a his tag. The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  The expression of Indoleamine 2, 3-dioxygenase is upregulated in response to IFN gamma treatment (PMID:30809284, 34735466).
  In Western blot, anti-H3 antibody (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) loading control staining at 1/100000 dilution.
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847) at 1/1000 dilution

  Lane 1: Untreated C2C12 (mouse myoblast) whole cell lysate at 20 µg

  Lane 2: C2C12 treated with /ml IFN gamma for 24 h, whole cell lysate at 20 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

  Observed band size: 45 kDa

  Exposure time: 180s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: no staining on rat liver. The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  The expression of Indoleamine 2, 3-dioxygenase is upregulated in response to IFN gamma treatment (PMID:30809284, 34735466).
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847) at 1/1000 dilution

  Lane 1: Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 2: RAW264.7 treated with /ml IFN gamma for 24 h, whole cell lysate at 20 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

  Observed band size: 45 kDa

  Exposure time: 180s

• 

  Flow Cytometry (Intracellular) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype control (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with an IDO1 expression vector containing a his tag (Middle) / HEK-293T transfected with IDO2 expression vector containing a his tag (Right) cells labelling Indoleamine 2, 3-dioxygenase with ab311847 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: no staining on mouse liver (PMID: 19741271). The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded (A) C2C12 (mouse myo tissue labelling Indoleamine 2, 3-dioxygenase with ab311847 at 1/500 (1.016 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) C2C12 treated with 50uM IFN-gamma for 24 hours cell pellet (treated). No staining on (B) C2C12 cell pellet (untreated). The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded Rat epididymis tissue labeling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat epididymis. The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded Mouse small intestine tissue labelling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse small intestinal secretory epithelial cells (PMID: 31325428). The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Flow Cytometry (Intracellular) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype control (Left) / C2C12 (mouse myoblast) treated with 50ng/ml IFN-γ for 24h (Middle) / Untreated C2C12 (Right) cells labelling Indoleamine 2, 3-dioxygenase with ab311847 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Immunohistochemical analysis of paraffin-embedded Mouse epididymis tissue labeling Indoleamine 2, 3-dioxygenase with ab311847 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse epididymis (PMID:19741271). The section was incubated with ab311847 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.
  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Heat-mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunoprecipitation - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847)

  Indoleamine 2, 3-dioxygenase was immunoprecipitated from 0.35 mg C2C12 (mouse myoblast) treated with 50 ng/ml IFN gamma for 24 h, whole cell lysate with ab311847 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab311847 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  Lane 1: C2C12 (mouse myoblast) treated with 50 ng/ml IFN gamma for 24 h, whole cell lysate
  Lane 2: C2C12 (mouse myoblast) treated with 50 ng/ml IFN gamma for 24 h, whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab311847 in C2C12 treated with 50 ng/ml IFN gamma for 24 h whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 180 seconds

  All lanes: Immunoprecipitation - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR28349-89] (ab311847) at 1/30 dilution

  All lanes: C2C12 (mouse myoblast) treated with 50 ng/ml IFN gamma for 24 h, whole cell lysate at 20 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 45 kDa

  Exposure time: 180s