Anti-Interferon gamma/IFNG Antibody Picoband®

Western blot analysis of IFNG using anti-IFNG antibody (A00393-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: rat spleen tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse thymus tissue 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-IFNG antigen affinity purified polyclonal antibody (Catalog # A00393-3) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IFNG at approximately 19 kDa. The expected band size for IFNG is at 19 kDa.

IHC analysis of IFNG using anti-IFNG antibody (A00393-3).
IFNG was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IFNG Antibody (A00393-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of IFNG using anti-IFNG antibody (A00393-3).
IFNG was detected in a paraffin-embedded section of mouse colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IFNG Antibody (A00393-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Th1/Th2 imbalance and Th17 immune response. ( A ) Immunohistochemistry and average optical density for IFN-γ, F DBP = 5.596 (p = 0.002), F TG = 142.67 (p = 0.000), F DBP*TG = 1.114 (p = 0.351). ( B ) Immunohistochemistry and average optical density for IL-4, F DBP = 0.537 (p = 0.659), F TG = 1.419 (p = 0.24), F DBP*TG = 0.168 (p = 0.917). ( C ) Immunohistochemistry and average optical density for IL-17 F DBP = 6.966 (p = 0.001), F TG = 93.46 (p = 0.000), F DBP*TG = 4.136 (p = 0.011). Magnification = ×40. *p < 0.05, **p < 0.01, compared with the saline group; # p < 0.05, ## p < 0.01, compared with the TG group; & p < 0.05, && p < 0.01, compared with the DBP50 + TG group.
Index in PubMed under a CC BY license. PMID: 29133889

AKT1 over-expression promoted cytokines release by MSCs. a The mRNA expression of IL-4, IL-10, PTGES2, HGF and VEGF in Null-MSCs and AKT1-MSCs were measured by qRT-PCR under routine culture condition and IFN-γ stimulation condition. The gene transcript levels were normalized to those of GAPDH and set to 1 in Null-MSCs untreated group. b ELISA analysis for detecting IL-10, PGE2, VEGF and HGF protein expression in MSCs culture supernatant after IFN-γ stimulation for 24h. The data are presented as the means ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001
Index in PubMed under a CC BY license. PMID: 31889917

Cytoprotective effect of AKT1 over-expression on MSCs. a Representative flow cytometry scatter plots depicting the percentage of apoptotic cells in Null-MSCs and AKT1-MSCs group under routine culture condition and IFN-γ stimulation condition. b The percentage of Annexin V-positive cells in each group is indicated in a bar chart ( left ). The ratio of Annexin V-positive cells under IFN-γ stimulation condition relative to untreated controls is indicated in a bar chart ( right ). The data are presented as the means ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. c Liver fluorescence images on day1, day7 and day14 after transplanting Null-MSCs or AKT1-MSCs into mice with or without Con A administration. d Relative quantification analysis of Null-MSCs and AKT-MSCs on day7 and day14 in ConA administration group. Fluorescence signals were quantified by ROI analysis using Living Image software 4.2 and normalized to those of Day1 in each group
Index in PubMed under a CC BY license. PMID: 31889917

Effects of anti-PD-1 and RSV on T cells and IFN-γ in testicular tissue. Percentages of testicular T cells, including CD4 + T cells (A) and CD8 + T cells (B) , were analyzed by flow cytometry in tumor-bearing mice after the final treatment (n = 4 at least, in each group). The mRNA level of IFN-γ in the testis was analyzed by qRT–qPCR (C) and the protein level of IFN-γ was analyzed by western blot (D) (n = 3 per group). Data are presented as means ± SEM. *P < 0.05 vs. Control group; #P < 0.05 vs. ICI group.
Index in PubMed under a CC BY license. PMID: 40145083