Anti-NCOA4 Antibody Picoband®

Western blot analysis of NCOA4 using anti-NCOA4 antibody (A04368-3).
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 293T whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human HL-60 whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human Daudi whole cell lysates,
Lane 7: human A431 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-NCOA4 antigen affinity purified polyclonal antibody (Catalog # A04368-3) 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 NCOA4 at approximately 70 kDa. The expected band size for NCOA4 is at 70 kDa.

IRN inhibits ferroptosis in db/db mice. (A–C) Examination of iron content, MDA levels, and GSH levels in mouse renal tissues. (D–I) Evaluation of SLC7A11, GPX4, TFR-1, FTH-1, and NCOA4 protein expression in mouse kidney tissues through western blotting and densitometric analysis of the bands. Results are presented as the mean ± SD of 6 mice. ***p < 0.001 versus db/m; ###p < 0.001 versus db/db.
Index in NEFROLOGIA under a CC BY license. DOI: 10.1016/j.nefro.2025.501408

IRN curbs HG-induced HK-2 cell injury and ferroptosis. (A) Detection of the cytotoxicity of IRN against HK-2 cells through CCK-8 assay. *p < 0.05. (B) Assessment of HK-2 cell viability after treatment with normal glucose, normal glucose + mannitol, high glucose, or high glucose + IRN (5, 10, 25, or 50 μM) by CCK-8 assay. (C–E) Measurement of iron content, MDA levels, and GSH levels in HK-2 cells. (F–K) Estimation of SLC7A11, GPX4, TFR-1, FTH-1, and NCOA4 protein levels in HK-2 cells via western blotting and densitometric analysis of the bands. Results are presented as the mean ± SD of three independent experiments. ***p < 0.001 versus NG; #p < 0.05, ##p < 0.01, ###p < 0.001 versus HG.
Index in NEFROLOGIA under a CC BY license. DOI: 10.1016/j.nefro.2025.501408

Western blot analysis of NCOA4 using anti-NCOA4 antibody (A04368-3).
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: rat liver tissue lysates,
Lane 2: rat RH35 whole cell lysates,
Lane 3: mouse liver tissue lysates,
Lane 4: mouse HEPA1/6 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-NCOA4 antigen affinity purified polyclonal antibody (Catalog # A04368-3) 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 NCOA4 at approximately 70 kDa. The expected band size for NCOA4 is at 70 kDa.

Flow Cytometry analysis of HL-60 cells using anti-NCOA4 antibody (A04368-3).
Overlay histogram showing HL-60 cells stained with A04368-3 (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-NCOA4 Antibody (A04368-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.