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[IF=7.376] Yunjia Li. et al. Indoleamine 2, 3-dioxygenase 1 aggravates acetaminophen-induced acute liver failure by triggering excess nitroxidative stress and iron accumulation. Free Radical Bio Med. 2021 Aug;172:578  IHC ;  Mouse.  

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[IF=1.96] Fu, Jingjing, et al. "Effect of bone marrow-derived CD11b+ F4/80+ immature dendritic cells on the balance between pro-inflammatory and anti-inflammatory cytokines in DBA/1 mice with collagen-induced arthritis." Inflammation Research (2014): 1-11.  Mouse.  

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[IF=0.7] Jiang, N., et al. "Expression of indoleamine 2, 3-dioxygenase in a murine model of Aspergillus fumigatus keratitis."International Journal of Ophthalmology 9.4 (2016).  IF(IHC-P) ;  Mouse.  

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Sample:
Lane 1: Mouse Epididymis tissue lysates
Lane 2: Human Hela cell lysates
Lane 3: Human Molt-4 cell lysates
Primary: Anti-IDO (bs-15493R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 45 kD
Observed band size: 48 kD

Tissue/cell: human lung carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-IDO Polyclonal Antibody, Unconjugated(bs-15493R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, AF647 conjugated(bs-0295G-A647)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei

Blank control: Mouse spleen.
Primary Antibody (green line): Rabbit Anti-IDO antibody (bs-15493R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-APC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control(blue):Hela (fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody:Rabbit Anti- IDO antibody(bs-15493R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.