AB252586

Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture)

RabMAb
Recombinant
KO Validated
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Rabbit Recombinant Monoclonal Indoleamine 2, 3-dioxygenase antibody. Carrier free. Suitable for WB, sELISA and reacts with Human samples.

View Alternative Names

IDO, INDO, IDO1, IDO-1

2 Images

Lab

Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture) (AB252586)

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.

All lanes:

Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (<a href='/en-us/products/primary-antibodies/indoleamine-2-3-dioxygenase-antibody-epr20374-ab211017'>ab211017</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate at 20 µg

Lane 2:

Wild-type A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate at 20 µg

Lane 2:

Western blot - Human IDO1 (Indoleamine 2, 3-dioxygenase) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-ido1-indoleamine-2-3-dioxygenase-knockout-a549-cell-line-ab266949'>ab266949</a>)

Lane 3:

IDO1 knockout A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate at 20 µg

Lane 4:

IDO1 knockout A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate at 20 µg

Lane 5:

SK-OV-3 cell lysate at 20 µg

Lane 6:

MCF7 cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 40 kDa

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Sandwich ELISA - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture) (AB252586)

sELISA

Supplier Data

Sandwich ELISA - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture) (AB252586)

Representative standard curve from corresponding SimpleStep ELISA® Kit (ab245710).

Unconjugated

Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]
Carrier free

Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - Low endotoxin, Azide free
617 Alexa Fluor® 594

Alexa Fluor® 594 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]
665 Alexa Fluor® 647

Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]
565 Alexa Fluor® 555

Alexa Fluor® 555 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]
Carrier free

Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free
519 Alexa Fluor® 488

Alexa Fluor® 488 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR20374

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

sELISA, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "sELISA" : {"fullname" : "Sandwich ELISA", "shortname":"sELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "sELISA-species-checked": "guaranteed", "sELISA-species-dilution-info": "", "sELISA-species-notes": "<p></p>" } } }

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Product details

ab252586 is a BSA and Azide Free antibody supplied in an unconjugated format and it is suitable for sandwich ELISAs to quantify Human IDO. The recommended pair for sandwich ELISA is:
Capture: ab252586, Human IDO Capture Antibody (unconjugated)
Detector: ab252645, Human IDO Detector Antibody (unconjugated)
The reference range value is 78.125 - 5000 pg/ml.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Sandwich ELISA
The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Indoleamine 2 3-dioxygenase (IDO) also known as IDO1 is an enzyme involved in the catabolism of tryptophan to kynurenine. It weighs approximately 45 kDa and it is an important target in immunoregulatory processes. IDO is highly expressed in antigen-presenting cells such as dendritic cells and macrophages as well as in various tumor cells where it helps modulate immune responses. This enzyme can also be measured by using assays like IDO ELISA.

Biological function summary

IDO plays a role in immune response regulation by degrading tryptophan an essential amino acid needed for T-cell proliferation. The depletion of tryptophan and the accumulation of its metabolites such as kynurenine cause immunosuppressive effects within the tissue microenvironment. While IDO functions mostly as a standalone enzyme its activity influences the cellular surroundings by altering the balance of local immune reactions.

Pathways

IDO integrates into the tryptophan metabolism pathway and is important in the kynurenine pathway. Through this pathway it maintains immune homeostasis and cell defense mechanisms. Proteins like kynureninase and kynurenine 3-monooxygenase also participate in the same metabolic processes which further extend the effects of tryptophan metabolism on immune cell behavior.

IDO is linked with cancer and chronic inflammatory conditions. In cancer elevated IDO expression suppresses anti-tumor immunity aiding tumor cells to evade immune surveillance. It also plays a role in autoimmune disorders by regulating excessive immune activity. During cancer progression IDO often works in conjunction with immune checkpoint proteins like PD-L1 which further enhances its immunosuppressive capabilities within the tumor microenvironment.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Catalyzes the first and rate limiting step of the catabolism of the essential amino acid tryptophan along the kynurenine pathway (PubMed : 17671174). Involved in the peripheral immune tolerance, contributing to maintain homeostasis by preventing autoimmunity or immunopathology that would result from uncontrolled and overreacting immune responses (PubMed : 25691885). Tryptophan shortage inhibits T lymphocytes division and accumulation of tryptophan catabolites induces T-cell apoptosis and differentiation of regulatory T-cells (PubMed : 25691885). Acts as a suppressor of anti-tumor immunity (PubMed : 14502282, PubMed : 23103127, PubMed : 25157255, PubMed : 25691885). Limits the growth of intracellular pathogens by depriving tryptophan (PubMed : 25691885). Protects the fetus from maternal immune rejection (PubMed : 25691885).

See full target information IDO1

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture)

Western blot - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture) (AB252586)

Sandwich ELISA - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (Capture) (AB252586)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

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Assay Kits

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Product promise