IDO Recombinant Rabbit Monoclonal Antibody [PSH12-31]
Catalog# HA723456
IDO Recombinant Rabbit Monoclonal Antibody [PSH12-31]
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WB
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IF-Cell
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IHC-P
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FC
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IP
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Human
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unconjugated
Overview
Product Name
IDO Recombinant Rabbit Monoclonal Antibody [PSH12-31]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human IDO aa 1-403.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 45 kDa
Positive Control
HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate, A549 treated with 50ng/mL hIFN-γ for 24 hours cell lysate, HeLa cells treated with 50ng/mL hIFN-γ for 16 hours, human colon adenocarcinoma tissue, human placenta tissue, human spleen tissue.
Conjugation
unconjugated
Clone Number
PSH12-31
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
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.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:100
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IHC-P
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1:1,000
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FC
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1:1,000
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IP
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1-2μg/sample
Target
Function
Indoleamine-pyrrole 2,3-dioxygenase (IDO or INDO EC 1.13.11.52) is a heme-containing enzyme physiologically expressed in a number of tissues and cells, such as the small intestine, lungs, female genital tract or placenta. In humans is encoded by the IDO1 gene. IDO is involved in tryptophan metabolism. It is one of three enzymes that catalyze the first and rate-limiting step in the kynurenine pathway, the O2-dependent oxidation of L-tryptophan to N-formylkynurenine, the others being indolamine-2,3-dioxygenase 2 (IDO2) and tryptophan 2,3-dioxygenase (TDO). IDO is an important part of the immune system and plays a part in natural defense against various pathogens. It is produced by the cells in response to inflammation and has an immunosuppressive function because of its ability to limit T-cell function and engage mechanisms of immune tolerance. Emerging evidence suggests that IDO becomes activated during tumor development, helping malignant cells escape eradication by the immune system. Expression of IDO has been described in a number of types of cancer, such as acute myeloid leukemia, ovarian cancer or colorectal cancer. IDO is part of the malignant transformation process and plays a key role in suppressing the anti-tumor immune response in the body, so inhibiting it could increase the effect of chemotherapy as well as other immunotherapeutic protocols. Furthermore, there is data implicating a role for IDO1 in the modulation of vascular tone in conditions of inflammation via a novel pathway involving singlet oxygen.
Background References
1. Salminen A. Role of indoleamine 2,3-dioxygenase 1 (IDO1) and kynurenine pathway in the regulation of the aging process. Ageing Res Rev. 2022 Mar
2. Shi D et al. USP14 promotes tryptophan metabolism and immune suppression by stabilizing IDO1 in colorectal cancer. Nat Commun. 2022 Sep
Subcellular Location
Cytoplasm, cytosol.
UNIPROT
Synonyms
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of IDO on different lysates with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of HeLa cells untreated / treated with 50ng/mL hIFN-γ for 16 hours labeling IDO with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) 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 (HA601187, 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. -
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma tissue with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) 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. -
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) 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. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) 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. -
☑ Relative expression (RE)
Immunohistochemical analysis of paraffin-embedded human liver tissue (negative) with Rabbit anti-IDO antibody (HA723456) 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 (HA723456) 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. -
☑ Cell treatment (CT)
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 (HA723456, 1/1,000) (red) 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
☑ Cell treatment (CT)
IDO was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate with HA723456 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723456 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: HA723456 IP in HeLa treated with 50ng/mL hIFN-γ for 16 hours cell lysate
Lane 3: Rabbit IgG instead of HA723456 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
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"