Recombinant Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (ab271990)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20374] to Indoleamine 2, 3-dioxygenase - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free
See all Indoleamine 2, 3-dioxygenase primary antibodies -
Description
Rabbit monoclonal [EPR20374] to Indoleamine 2, 3-dioxygenase - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type A549 Treated IFN gamma, Human ovary cancer, placenta and tonsil lysates; SK-OV-3 whole cell lysate; HeLa whole cell lysate treated with 50ng/ml Interferon-gamma (IFN-gamma) for 16 hours. IHC-P: Human spleen, tonsil, placenta and endometrium cancer tissues. ICC/IF: HeLa cells treated with IFN-gamma (50 ng/ml) for 16 hours. Flow Cyt (intra): HeLa cells treated with IFN-gamma (50 ng/ml) for 16 hours. IP: HeLa whole cell lysate treated with 50ng/ml IFN-gamma for 16h.
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General notes
ab271990 is the carrier-free version of ab211017.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20374 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271990 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 45 kDa.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 45 kDa. |
Target
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Function
Catalyzes the cleavage of the pyrrol ring of tryptophan and incorporates both atoms of a molecule of oxygen. -
Pathway
Amino-acid degradation; L-tryptophan degradation via kynurenine pathway; L-kynurenine from L-tryptophan: step 1/2. -
Sequence similarities
Belongs to the indoleamine 2,3-dioxygenase family. - Information by UniProt
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Database links
- Entrez Gene: 3620 Human
- Omim: 147435 Human
- SwissProt: P14902 Human
- Unigene: 840 Human
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Alternative names
- 3-dioxygenase antibody
- I23O1_HUMAN antibody
- IDO 1 antibody
see all
Images
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All lanes : Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution
Lane 1 : Wild-type A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate
Lane 2 : Wild-type A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate
Lane 3 : IDO1 knockout A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate
Lane 4 : IDO1 knockout A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate
Lane 5 : SK-OV-3 cell lysate
Lane 6 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab211017).
Lanes 1 - 6: Merged signal (red and green). Green - ab211017 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab211017 was shown to react with Indoleamine 2, 3-dioxygenase in treated wild-type A549 cells in Western blot with no signal observed in treated IDO1 knockout cell line ab266949 (IDO1 knockout cell lysate ab256948). Wild-type A549 and IDO1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab211017 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on dendritic cells of human spleen is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017). -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on dendritic cells of human tonsil is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017). -
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on endothelial cells of human placenta is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017). -
Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on human endometrium cancer is observed (PMID: 26155395).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017). -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-γ for 16 hours or untreated, labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The signal increased after treatment with IFN-γ (50 ng/ml) for 16 hours on HeLa cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017). -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-gamma for 16h (red) or untreated (green), labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017).
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Indoleamine 2, 3-dioxygenase was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 50ng/ml IFN-γ for 16h with ab211017 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab211017 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate 10 µg (Input).
Lane 2: ab211017 IP in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211017 in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab211017).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab271990 has not yet been referenced specifically in any publications.