IDO1 Rabbit Polyclonal Antibody

• Catalog Number: orb500950
• Category: Antibodies
• Description: IDO1 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: WB
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Rabbit, Rat, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human IDO (101-200/403aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 45 kDa
• Target: IDO1
• UniProt ID: P14902
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Indoleamine 2, 3-dioxygenase; 3-dioxygenase; I23O1
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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 (orb500950), dilution: 1 µg 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 1X PBS containing 0.5% BSA.

Blank control: Mouse spleen. Primary Antibody (green line): Rabbit Anti-IDO antibody (orb500950), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-APC, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min 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 20000 events was performed.

Sample: Lane 1: Mouse Epididymis tissue lysates, Lane 2: Human Hela cell lysates, Lane 3: Human Molt-4 cell lysates, Primary: Anti-IDO (orb500950) 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 15 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-IDO Polyclonal Antibody, Unconjugated (orb500950) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, AF647 conjugated used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.