WB
Western blot analysis of IDO on different lysates with Rabbit anti-IDO antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Lane 3: A549 cell lysate Lane 4: A549 treated with 50ng/mL hIFN-γ for 24 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 45 kDa Observed band size: 45 kDa Exposure time: 2 seconds; ECL: K1801; 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 at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HeLa cells untreated / treated with 50ng/mL hIFN-γ for 16 hours labeling IDO with Rabbit anti-IDO antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L 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 was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-IDO antibody at 1/1,000 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 at 1/1,000 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.FC
Flow cytometric analysis of HeLa cells untreated (left) / treated with 50ng/mL hIFN-γ for 16 hours (right) labeling IDO. Cells were fixed and permeabilized. Then stained with the primary antibody compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
IDO was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate (input) Lane 2: IP in HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Lane 3: Rabbit IgG instead of in HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 6 seconds; ECL: K1801| Product Name | IDO Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human IDO aa 1-403. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHC-PIPWB |
| WB:1:5,000 IHC:1:1,000 FC:1:1,000 IP:1-2μg/sample IF: 1:100 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | IDO1 |
|---|---|
| Gene Synonyms | IDO INDO IDO-1 |
| Gene Full Name | indoleamine 2,3-dioxygenase 1 |
| Gene Summary | This gene encodes indoleamine 2,3-dioxygenase (IDO) - a heme enzyme that catalyzes the first and rate-limiting step in tryptophan catabolism to N-formyl-kynurenine. This enzyme acts on multiple tryptophan substrates including D-tryptophan, L-tryptophan, 5-hydroxy-tryptophan, tryptamine, and serotonin. This enzyme is thought to play a role in a variety of pathophysiological processes such as antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Through its expression in dendritic cells, monocytes, and macrophages this enzyme modulates T-cell behavior by its peri-cellular catabolization of the essential amino acid tryptophan.[provided by RefSeq, Feb 2011] |
| Alternative Names | 3-dioxygenase antibody I23O1_HUMAN antibody IDO 1 antibody IDO antibody IDO-1 antibody IDO1 antibody INDO antibody indolamine 2 3 dioxygenase antibody Indole 2 3 dioxygenase antibody indoleamine 2 3 dioxygenase 1 antibody 3-dioxygenase antibody I23O1_HUMAN antibody IDO 1 antibody IDO antibody IDO-1 antibody IDO1 antibody INDO antibody indolamine 2 3 dioxygenase antibody Indole 2 3 dioxygenase antibody indoleamine 2 3 dioxygenase 1 antibody indoleamine 2 3 dioxygenase antibody Indoleamine 2 3-dioxygenase 1 antibody Indoleamine pyrrole 2 3 dioxygenase antibody Indoleamine-pyrrole 2 antibody |
| Molecular Weight(MW) | 45 kDa |
| Cellular Localization | Cytoplasm |
WB
Western blot analysis of IDO on different lysates with Rabbit anti-IDO antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Lane 3: A549 cell lysate Lane 4: A549 treated with 50ng/mL hIFN-γ for 24 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 45 kDa Observed band size: 45 kDa Exposure time: 2 seconds; ECL: K1801; 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 at 1/5,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells untreated / treated with 50ng/mL hIFN-γ for 16 hours labeling IDO with Rabbit anti-IDO antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L 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 was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-IDO antibody at 1/1,000 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 at 1/1,000 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.
FC
Flow cytometric analysis of HeLa cells untreated (left) / treated with 50ng/mL hIFN-γ for 16 hours (right) labeling IDO. Cells were fixed and permeabilized. Then stained with the primary antibody compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
IDO was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate with at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate (input) Lane 2: IP in HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Lane 3: Rabbit IgG instead of in HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 6 seconds; ECL: K1801| Application Notes | WB:1:5,000 IHC:1:1,000 FC:1:1,000 IP:1-2μg/sample IF: 1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
