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Indoleamine 2,3-Dioxygenase 1 Peptide 4

Antibody: CPTC-IDO1-6
Background

Catalog Number:

CPTC-IDO1-6

Target Antigen:

Indoleamine 2,3-Dioxygenase 1 Peptide 4

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

04/20/2023

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous

Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image The affinity and binding kinetics of CPTC-IDO1-6 antibody and BSA-conjugated peptide “GTGGTDLMNFLK" were measured using biolayer interferometry. BSA-conjugated peptide was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-IDO1-6 antibody, 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. Buffer only and biosensors immobilized without BSA-conjugated peptide were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model. Click image to enlarge

CPTC-IDO1-6 Affinity and Kinetics (Bio-Layer Interferometry)

Result: Positive

The affinity and binding kinetics of CPTC-IDO1-6 antibody and BSA-conjugated peptide “GTGGTDLMNFLK" were measured using biolayer interferometry. BSA-conjugated peptide was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-IDO1-6 antibody, 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. Buffer only and biosensors immobilized without BSA-conjugated peptide were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image The affinity and binding kinetics of CPTC-IDO1-6 antibody and BSA-conjugated peptide, “GTGGTDLMNFLK”, were measured using surface plasmon resonance. Peptide was amine coupled onto a Series S CM5 biosensor chip. Antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model. Click image to enlarge

CPTC-IDO1-6 Affinity and Kinetics (Surface Plasmon Resonance) 

Result: Positive

The affinity and binding kinetics of CPTC-IDO1-6 antibody and BSA-conjugated peptide, “GTGGTDLMNFLK”, were measured using surface plasmon resonance. Peptide was amine coupled onto a Series S CM5 biosensor chip. Antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM, 0.25 nM, and 0.0625 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent fitting model.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM using CPTC-IDO1-6 as capture antibody against the synthetic peptide GTGGTDLMNFLK. Antibody CPTC-IDO1-6 captures the synthetic peptide. Click image to enlarge

CPTC-IDO1-6 iMRM (synthetic peptide)

Result: Positive

Immuno-MRM using CPTC-IDO1-6 as capture antibody against the synthetic peptide GTGGTDLMNFLK. Antibody CPTC-IDO1-6 captures the synthetic peptide.


Characterization SOP Files

  Immunofluorescence

CPTC-IDO1-6 Immunofluorescence

Result: Negative

Immunofluorescence staining using CPTC-IDO1-6 as primary antibody against MCF7 cells shows no localization of IDO1 protein.


Characterization SOP Files

  Indirect ELISA
Click to enlarge image Indirect ELISA using CPTC-IDO1-6 as primary antibody against BSA conjugated peptide, "GTGGTDLMNFLK", coated on the plate and detected using HRP conjugated goat anti-rabbit secondary antibody and TMB. Click image to enlarge

CPTC-IDO1-6 Indirect ELISA

Result: Positive

Indirect ELISA using CPTC-IDO1-6 as primary antibody against BSA conjugated peptide, "GTGGTDLMNFLK", coated on the plate and detected using HRP conjugated goat anti-rabbit secondary antibody and TMB.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-IDO1-6 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Western blot using CPTC-IDO1-6 as primary antibody against human indoleamine 2,3-dioxygenase 1 (IDO1) recombinant protein. The expected molecular weight is 45.1 kDa. Click image to enlarge

CPTC-IDO1-6 Western Blot (Recombinant Protein)

Result: Presumed Positive (with additional bands)

Western blot using CPTC-IDO1-6 as primary antibody against human indoleamine 2,3-dioxygenase 1 (IDO1) recombinant protein. The expected molecular weight is 45.1 kDa.


Characterization SOP Files

  Western Blot - Recombinant Protein or Peptide
Click to enlarge image Automated western blot using CPTC-IDO1-6 as primary antibody against recombinant IDO1 protein. Protein molecular weight is about 45 KDa. The antibody presumably recognizes recombinant IDO1. Click image to enlarge

CPTC-IDO1-6 Automated Western Blot (Recombinant Protein)

Result: Presumed Positive (with additional bands)

Automated western blot using CPTC-IDO1-6 as primary antibody against recombinant IDO1 protein. Protein molecular weight is about 45 KDa. The antibody presumably recognizes recombinant IDO1.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-IDO1-6 as primary antibody against whole cell lysates CCRF-CEM (lane 2), HeLa (lane 3), Jurkat (lane 4), K-562 (lane 5), and MCF7 (lane 6). The expected molecular weight is 45.3 kDa. All cell lines are positive. Click image to enlarge

CPTC-IDO1-6 Western Blot (Cell Lysates)

Result: Positive

Western blot using CPTC-IDO1-6 as primary antibody against whole cell lysates CCRF-CEM (lane 2), HeLa (lane 3), Jurkat (lane 4), K-562 (lane 5), and MCF7 (lane 6). The expected molecular weight is 45.3 kDa. All cell lines are positive.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated western blot using CPTC-IDO1-6 as primary antibody against whole lysates of cell lines CCRF-CEM, HeLa, Jurkat, K-562 and MCF7. Protein molecular weight is about 45 KDa. The antibody cannot recognize IDO1 in the tested lysates. Loading controls were run with anti-GAPDH antibody. Click image to enlarge

CPTC-IDO1-6 Automated Western Blot (Cell Lysates)

Result: Negative

Automated western blot using CPTC-IDO1-6 as primary antibody against whole lysates of cell lines CCRF-CEM, HeLa, Jurkat, K-562 and MCF7. Protein molecular weight is about 45 KDa. The antibody cannot recognize IDO1 in the tested lysates. Loading controls were run with anti-GAPDH antibody.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate

CPTC-IDO1-6 Single Cell Western Blot (Cell Lysate)

Result: Negative

Single cell western blot using CPTC-IDO1-6 as a primary antibody against MCF7 cell lysate. IDO1 protein expression was not detected.


Characterization SOP Files

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Antigen: Indoleamine 2,3-Dioxygenase 1 Peptide 4
Background

NCI Identification Number:

00524

Antigen Name:

Indoleamine 2,3-Dioxygenase 1 Peptide 4

CPTC Name:

CPTC-IDO1 Peptide 4

Aliases:

Indoleamine 2,3-Dioxygenase 1; INDO; IDO; Indoleamine-Pyrrole 2,3-Dioxygenase; IDO-1; Indoleamine-Pyrrole 2,3 Dioxygenase; Indolamine 2,3 Dioxygenase; Indole 2,3-Dioxygenase; EC 1.13.11.52

Function:

This gene encodes indoleamine 2,3-dioxygenase (IDO) - a heme enzyme that catalyzes the first and rate-limiting step in tryptophan catabolism to N-formyl-kynurenine. This enzyme acts on multiple tryptophan substrates including D-tryptophan, L-tryptophan, 5-hydroxy-tryptophan, tryptamine, and serotonin. This enzyme is thought to play a role in a variety of pathophysiological processes such as antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Through its expression in dendritic cells, monocytes, and macrophages this enzyme modulates T-cell behavior by its peri-cellular catabolization of the essential amino acid tryptophan.IDO1 (Indoleamine 2,3-Dioxygenase 1) is a Protein Coding gene. Diseases associated with IDO1 include Listeriosis and Trypanosomiasis. Among its related pathways are Histidine, lysine, phenylalanine, tyrosine, proline and tryptophan catabolism and superpathway of tryptophan utilization. Gene Ontology (GO) annotations related to this gene include heme binding and tryptophan 2,3-dioxygenase activity.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:23103127, PubMed:25157255, PubMed:14502282, PubMed:25691885). Limits the growth of intracellular pathogens by depriving tryptophan (PubMed:25691885). Protects the fetus from maternal immune rejection (PubMed:25691885).

Chromosomal Localization:

8p11.21

Accession Number:

NP_002155.1

UniProt Accession Number:

P14902

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

GTGGTDLMNFLK

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

1320

Last Updated:

06/09/2022

Links

Characterization Data

SOPs

No SOPs available.

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