IDO Recombinant Rabbit Monoclonal Antibody [PD00-62]
cat.: HA721331
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: IHC-P, IF-Cell, WB
Clonality: Monoclonal
Clone number: PD00-62
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 45 kDa
Isotype: IgG
Immunogen: Synthetic peptide derived from the C-terminus of human IDO-1 protein.
Positive control: Human endometrial carcinoma tissue, human lymph nodes tissue, human placenta tissue, HeLa cells treated with 50 ng/mL IFN-gamma for 16 hours, A549 treated with 50ng/mL IFN-gamma for 24 hours cell lysate.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  IHC-P
  IF-Cell
  WB

1:200
1:100
1:1,000
Uniprot #: SwissProt: P14902 Human
Alternative names: 3-dioxygenase I23O1_HUMAN IDO 1 IDO IDO-1 IDO1 INDO indolamine 2,3 dioxygenase Indole 2 3 dioxygenase indoleamine 2 3 dioxygenase 1 indoleamine 2 3 dioxygenase Indoleamine 2,3-dioxygenase 1 Indoleamine pyrrole 2 3 dioxygenase Indoleamine-pyrrole 2
Images
HA721331_1.jpg Fig1: Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue with Rabbit anti-IDO antibody (HA721331) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721331) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721331_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-IDO antibody (HA721331) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721331) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721331_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-IDO antibody (HA721331) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721331) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721331_4.jpg Fig4: Immunocytochemistry analysis of HeLa cells treated with or without 50 ng/mL IFN-gamma for 16 hours labeling IDO with Rabbit anti-IDO antibody (HA721331) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IDO antibody (HA721331) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721331_5.jpg Fig5: Western blot analysis of IDO on different lysates with Rabbit anti-IDO antibody (HA721331) at 1/1,000 dilution.

Lane 1: A549 cell lysate
Lane 2: A549 treated with 50ng/mL IFN-gamma for 24 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 40 kDa

Exposure time: 17 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721331) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.