14-9750-82

antibody from Invitrogen Antibodies

Targeting: IDO1 IDO, INDO

Provider product page for 14-9750-82

Western blot

Immunohistochemistry

Antibody data

Antibody Data
Antigen structure
References [6]
Comments [0]
Validations
Western blot [2]
Immunohistochemistry [1]

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Product number: 14-9750-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: IDO Monoclonal Antibody (V1NC3IDO), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The monoclonal antibody V1NC3IDO recognizes human IDO. IDO (indoleamine-2,3-dioxygenase, INDO, IDO1) is an intracellular enzyme that catalyzes the degradation of tryptophan to kynurenines. IDO is expressed in a wide variety of tissues and cells, including macrophages, plasmacytoid dendritic cells, and several cell lines. IDO can be induced in many different cell types by IFN gamma or other inflammatory stimuli. Expression of IDO in antigen presenting cells and tumors is thought to mediate immune suppression through depletion of this essential amino acid and/or by the creation of tryptophan metabolites that cause apoptosis of T cells and induction of regulatory T cells. This V1NC3IDO antibody reacts with IDO1 and does not cross react with IDO2. Applications Reported: This V1NC3IDO antibody has been reported for use in immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, and microscopy. Applications Tested: This V1NC3IDO antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded human tissue using low pH antigen retrieval and can be used at less than or equal to 0.5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: V1NC3IDO
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: 4° C

IDO1 protein structure - 14-9750-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of IDO1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR864924_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of IDO1 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa wild type treated with 50 ng/mL IFN gamma for 24hrs (Lane 2), HeLa Cas9 (Lane 3), HeLa Cas9 treated with 50 ng/mL IFN gamma for 24hrs (Lane 4), HeLa IDO1 KO (Lane 5) and HeLa IDO1 KO treated with 50 ng/mL IFN gamma for 24hrs (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-IDO Monoclonal Antibody (V1NC3IDO), eBioscience™ (Product # 14-9750-82, 1;1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1;8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to IDO1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-IDO Monoclonal Antibody (V1NC3IDO), eBioscience™ (Product # 14-9750-82) and a 43 kDa band corresponding to Indoleamine 2,3-dioxygenase 1 was observed across SK-OV-3 and HEK-293 treated with IFN gamma. Whole cell extracts (30 µg lysate) of SK-O-V3 (Lane 1), HEK-293 (Lane 2), HEK-293 treated with 50 ng/ml of IFN gamma for 24 hours (Lane 3), MDA-MB-231 (Lane 4) and MCF7 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).