Anti-NOX2/gp91phox/CYBB Antibody Picoband®

Western blot analysis of CYBB using anti-CYBB antibody (PA1667).
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 Raji whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: rat RH35 whole cell lysates,
Lane 5: mouse RAW264.7 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-CYBB antigen affinity purified polyclonal antibody (Catalog # PA1667) 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 CYBB at approximately 65 kDa. The expected band size for CYBB is at 65 kDa.

IHC analysis of CYBB using anti-CYBB antibody (PA1667).
CYBB 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-CYBB Antibody (PA1667) 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.

Western blot analysis of NOX2/gp91phox/CYBB using anti-NOX2/gp91phox/CYBB antibody(PA1667).
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.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% milk for 1.5 hour at RT. The membrane was incubated with rabbit anti-NOX2/gp91phox/CYBB antibody(PA1667) at 1:1000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a Cell Signaling Technology's 7074s murine anti-rabbit conformation specific antibody at a dilution of 1:5000 for 1 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 NOX2/gp91phox/CYBB at approximately 65 kDa. The expected band size for NOX2/gp91phox/CYBB is at 65 kDa.

ASIV reduced ADR-induced oxidative stress. (A) Representative images of DHE staining and quantification of ROS production. DHE positive (red), nuclear stained with DAPI (blue). Serum GPx (B) , SOD (C) , and MDA (D) levels among the different groups ( n = 3). (E) Protein expression of oxidative stress-associated proteins, including NOX2, NOX4 in kidney tissue. (F,G) The expression of NOX2 and NOX4. Data were expressed as mean ± SD, n = 6. * p < 0.05, ** p < 0.01 vs. CON group. # p < 0.05, ## p < 0.01 vs. ADR group.
Index in PubMed under a CC BY license. PMID: 35370757