Thioredoxin (Trx) is a redox protein that is found in several species, such as bacteria, plants and mammals, and contains a conserved active site, consisting of Trp-Cys-Gly-Pro-Cys. Trx has several biological functions. It acts as a hydrogen donor for ribonucleotide reductase, which is critical for DNA synthesis, and modulates the DNA-binding activity of several transcription factors, including NFkB, AP-1, p53, TFIIIC and glucocorticoid receptor. Trx also stimulates cell growth, is an inhibitor of apoptosis and plays a role in the protection against oxidative stress. Drugs that inhibit Trx have antitumor activity, suggesting that Trx is involved in a variety of human diseases, including cancer. TrxR is a ubiquitously expressed flavoprotein that catalyzes the NADPH-dependent reduction of Trx as well as several other oxidized cellular components.
Background References
1. Masutani H. Thioredoxin-Interacting Protein in Cancer and Diabetes. Antioxid Redox Signal. 2022 May
2. Jastrząb A et al. Thioredoxin-dependent system. Application of inhibitors. J Enzyme Inhib Med Chem. 2021 Dec
Western blot analysis of Thioredoxin on different lysates with Rabbit anti-Thioredoxin antibody (HA724383) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate
Lane 2: Hep G2 (Human liver cancer cells) cell lysate
Lane 3: MCF7 (Human breast cancer cells) cell lysate
Lane 4: A549 (Human lung adenocarcinoma cells) cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 9 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724383, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 12 kDa
Observed band size: 12 kDa
Western blot analysis of Thioredoxin on different lysates with Rabbit anti-Thioredoxin antibody (HA724383) at 1/5,000 dilution.
Lane 1: COS-1 (African green monkey kidney fibroblasts) cell lysate
Lane 2: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 3: Neuro-2a (Mouse brain neuroblastoma cells) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 5: C2C12 (Mouse myoblasts) cell lysate
Lane 6: C6 (Rat glioma cells) cell lysate
Lane 7: PC-12 (Rat pheochromocytoma cells (undifferentiated)) cell lysate
Lane 8: Mouse kidney tissue lysate
Lane 9: Rat kidney tissue lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 9 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724383, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 12 kDa
Observed band size: 12 kDa
Immunoprecipitation (IP)
Thioredoxin was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724383 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724383 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724383 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724383 in HeLa cell lysate
Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724383, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 12 kDa
Observed band size: 12 kDa
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