Revision 7

#86630Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-IC, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

43

Source/Isotype:

Rabbit IgG

UniProt ID:

#P14902

Entrez-Gene Id:

3620

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
IHC Leica Bond 1:200 - 1:800
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #91473.

Specificity / Sensitivity

IDO (D5J4E) Rabbit mAb recognizes endogenous levels of total IDO (IDO-1, INDO) protein. The antibody does not cross-react with IDO-2 (INDOL1). Some nonspecific staining of normal breast epithelium has been observed.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant human IDO protein.

Background

INDO/IDO1/indoleamine 2,3-dioxygenase (IDO) is an IFN-γ-inducible enzyme that catalyzes the rate-limiting step of tryptophan degradation (1). IDO is upregulated in many tumors and in dendritic cells in tumor-draining lymph nodes. Elevated tryptophan catabolism in these cells leads to tryptophan starvation of T cells, limiting T cell proliferation and activation (2). Therefore, IDO is considered an immunosuppresive molecule, and research studies have shown that upregulation of IDO is a mechanism of cancer immune evasion (3). The gastrointestinal stromal tumor drug, imatinib, was found to act, in part, by reducing IDO expression, resulting in increased CD8+ T cell activation and induction of apoptosis in regulatory T cells (4). In addition to its enzymatic activity, IDO was recently shown to have signaling capability through an immunoreceptor tyrosine-based inhibitory motif (ITIM) that is phosphorylated by Fyn in response to TGF-β. This leads to recruitment of SHP-1 and activation of the noncanonical NF-κB pathway (5).

  1. Yasui, H. et al. (1986) Proc Natl Acad Sci U S A 83, 6622-6.
  2. Mellor, A.L. et al. (2003) Adv Exp Med Biol 527, 27-35.
  3. Prendergast, G.C. (2008) Oncogene 27, 3889-900.
  4. Balachandran, V.P. et al. (2011) Nat Med 17, 1094-100.
  5. Pallotta, M.T. et al. (2011) Nat Immunol 12, 870-8.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 7
#86630

IDO (D5J4E) Rabbit mAb

Western Blotting Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (50 ng/ml, 16 hr; +), using IDO (D5J4E) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Immunoprecipitation of IDO from extracts of HeLa cells treated with Human Interferon-γ (hIFN-γ) #8901 (50 ng/ml, 16 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IDO (D5J4E) Rabbit mAb. Western blot analysis was performed using IDO (D5J4E®) Rabbit mAb.
Immunohistochemistry Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using IDO (D5J4E) Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using IDO (D5J4E) Rabbit mAb.
Immunohistochemistry Image 2: IDO (D5J4E™) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using IDO (D5J4E) Rabbit mAb.
Immunohistochemistry Image 3: IDO (D5J4E™) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using IDO (D5J4E) Rabbit mAb.
Immunohistochemistry Image 4: IDO (D5J4E™) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Interferon-γ (hIFN-γ) #8901 (10 ng/ml, 16 hr; right), using IDO (D5J4E) Rabbit mAb.
Immunohistochemistry Image 5: IDO (D5J4E™) Rabbit mAb Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using IDO (D5J4E) rabbit mAb (yellow), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), PD-L2 (D7U8C) XP® rabbit mAb (magenta) #82723, Arginase-1 (D4E3M) XP® rabbit mAb #93668 (green), CD68 (D4B9C) XP® rabbit mAb #76437 (orange), and Pan-keratin (C11) mouse mAb #4545 (cyan).
Immunofluorescence Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Human Interferon-γ (hIFN-γ) #8901 (10 ng/ml, overnight; right), using IDO (D5J4E) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: IDO (D5J4E™) Rabbit mAb Expand Image
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with human interferon-γ (hIFN-γ) (10 ng/ml, 16 hr; green) using IDO (D5J4E) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.