Biolegend, 695002, Purified anti-IDO1 Antibody, 100μg

IDO1 is also known as Indolamine 2,3-dioxygenase, Indole 2,3-dioxygenase, and Indoleamine-pyrrole 2,3-dioxygenase. IDO1 is a ubiquitously expressed cytoplasmic protein with a predicted molecular weight of approximately 45 kD. It is one of the best known IFN-γ inducible genes. The product of IDO gene catalyzes the degradation of the essential amino acid L-tryptophan to N-formylkynurenine. IDO has been implicated to protect against intracellular and extracellular pathogens. It also has been shown to maintain the special immune suppressive status of immune-privileged sites such as the brain, eyes, kidney, and placenta.
100μg
Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Human IDO1 recombinant protein (154-403 a.a.) expressed in E. coli.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: WB - Quality tested SB - Community verified
Recommended Usage: Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 2.0 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.
Additional Product Notes: For the use of this antibody in spatial biology applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ can be found here.
Product Citations: Majumdar T, et al. 2019. Cell Death Dis. 10:161. PubMed
RRID: AB_2650744 (BioLegend Cat. No. 695001) AB_2650745 (BioLegend Cat. No. 695002)
Structure: 403 amino acids with a predicted molecular weight of 45 kD.
Distribution: Cytoplasm.
Function: Indoleamine 2,3-dioxygenase (IDO) is an IFN-γ inducible gene. It catalyzes the degradation of the essential amino acid L-tryptophan to N-formylkynurenine. IDO has been implicated to protect against intracellular and extracellular pathogens.
Interaction: This enzyme acts on multiple tryptophan substrates including D-tryptophan, L-tryptophan, 5-hydroxy-tryptophan, tryptamine, and serotonin.
Biology Area: Cancer Biomarkers, Cell Biology, Immunology, Innate Immunity
Antigen References: 1. Habara-Ohkubo A, et al. 1991. Gene. 105:221. 2. Munn DH, et al. 2002. Science. 297:1867. 3. Frumento G, et al. 2002. J. Exp. Med. 196:459. 4. Muller AJ, et al. 2005. Nature Med. 11:312. 5. Löb S, et al. 2009. Nature Rev. Cancer 9:445-52.
Gene ID: 15930
UniProt: View information about IDO1 on UniProt.org
Clone: W16073A
Regulatory Status: RUO
Other Names: Indolamine 2,3-dioxygenase, Indole 2,3-dioxygenase, Indoleamine-pyrrole 2,3-dioxygenase
Isotype: Rat IgG2a, κ
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.