Cysteine protease ATG4B is an enzyme that in humans is encoded by the ATG4B gene. Autophagy is the process by which endogenous proteins and damaged organelles are destroyed intracellularly. Autophagy is postulated to be essential for cell homeostasis and cell remodeling during differentiation, metamorphosis, non-apoptotic cell death, and aging. Reduced levels of autophagy have been described in some malignant tumors, and a role for autophagy in controlling the unregulated cell growth linked to cancer has been proposed. This gene encodes a member of the autophagin protein family. The encoded protein is also designated as a member of the C-54 family of cysteine proteases. Alternate transcriptional splice variants, encoding different isoforms, have been characterized. One main function of Atg4 is to cleave the pre-protein of Atg8, leading to the non-lipidated soluble (-I) form which can be processed further by Atg3, Atg7, Atg5-12 into the lipidated form (-II) anchored to the autophagic membrane.
Background References
1. Sun L et al. Deacetylation of ATG4B promotes autophagy initiation under starvation. Sci Adv. 2022 Aug
2. Yang G et al. Targeting Atg4B for cancer therapy: Chemical mediators. Eur J Med Chem. 2021 Jan
Western blot analysis of ATG4B on different lysates with Rabbit anti-ATG4B antibody (HA721829) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.
Lane 1: HeLa cell lysate Lane 2: Jurkat cell lysate Lane 3: 293T cell lysate Lane 4: MCF7 cell lysate Lane 5: K-562 cell lysate Lane 6: COS-1 cell lysate Lane 7: NIH/3T3 cell lysate Lane 8: C2C12 cell lysate Lane 9: PC-12 cell lysate Lane 10: Mouse brain tissue lysate Lane 11: Rat brain tissue lysate Lane 12: Mouse kidney tissue lysate Lane 13: Mouse colon tissue lysate
Lysates/proteins at 30 µg/Lane.
Predicted band size: 44 kDa Observed band size: 47 kDa
Exposure time: 42 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721829) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of ATG4B on different lysates with Rabbit anti-ATG4B antibody (HA721829) at 1/2,000 dilution.
Lane 1: 293T cell lysate (20 µg/Lane) Lane 2: MCF7 cell lysate (20 µg/Lane) Lane 3: COS-1 cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: PC-12 cell lysate (20 µg/Lane) Lane 6: Mouse brain tissue lysate (40 µg/Lane) Lane 7: Rat brain tissue lysate (40 µg/Lane)
Predicted band size: 44 kDa Observed band size: 47 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721829) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of ATG4B on different lysates with Rabbit anti-ATG4B antibody (HA721829) at 1/1,000 dilution.
Lane 1: 293T-si NT cell lysate Lane 2: 293T-si ATG4B cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 44 kDa Observed band size: 47 kDa
Exposure time: 51 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721829) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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