Anti-Indoleamine 2, 3-dioxygenase/IDO1 Antibody Picoband®

Western blot analysis of IDO1 using anti-IDO1 antibody (PB9603).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human K562 whole cell lysates,
Lane 3: human PC-3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-IDO1 antigen affinity purified polyclonal antibody (PB9603) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for IDO1 at approximately 45 kDa. The expected band size for IDO1 is at 45 kDa.

IHC analysis of IDO1 using anti-IDO1 antibody (PB9603).
IDO1 was detected in paraffin-embedded section of Human Lung Cancer Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-IDO1 Antibody (PB9603) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Flow Cytometry analysis of A431 cells using anti-IDO1 antibody (PB9603).
Overlay histogram showing A431 cells stained with PB9603 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IDO1 Antibody (PB9603,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of IDO1 using anti-IDO1 antibody (PB9603).
IDO1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-IDO1 Antibody (PB9603) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

The heterogeneity within the myeloid cells. a 4 subclusters of myeloid cells were identified by UMAP analysis. b Marker gene expression of each cell type. c Comparison of cell count proportion of each cell type between imatinib resistant and sensitive patients. d mIHC staining of panCK (red), LYZ (green), IDO1 (magenta) and DAPI in GIST TME.
Index in PubMed under a CC BY license. PMID: 38443340

Transcription characteristics and heterogeneity among the six malignant cell types. a UMAP analysis showing the six cell type of malignant cells. b Colored cells by each sample. c Bar plot showing the cell count proportion of each maligant subcluster in GIST. d GO enrichment analysis of marker genes form BASP1_fib (left), DUSP1_fib (middle), IDO1_fib (right).
Index in PubMed under a CC BY license. PMID: 38443340