One of the enzymes of the urea cycle, the metabolic pathway transforming neurotoxic amonia produced by protein catabolism into inocuous urea in the liver of ureotelic animals. Catalyzes the formation of arginosuccinate from aspartate, citrulline and ATP and together with ASL it is responsible for the biosynthesis of arginine in most body tissues.
Background References
1. Shaheen N et al. Characterization of human wild-type and mutant argininosuccinate synthetase proteins expressed in bacterial cells. Enzyme Protein 48:251-264 (1994).
2. Berning C et al. Investigation of citrullinemia type I variants by in vitro expression studies. Hum Mutat 29:1222-1227 (2008).
Sequence Similarity
Belongs to the argininosuccinate synthase family. Type 1 subfamily.
Tissue Specificity
Expressed in adult liver.
Post-translational Modification
Acetylated by CLOCK in a circadian manner which negatively regulates its enzyme activity. Deacetylated by histone deacetylases.
Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (EM1701-87) at 1/5,000 dilution.
Lane 1: MCF7 cell lysate, 15 µg/Lane Lane 2: Human liver tissue lysate, 30 µg/Lane Lane 3: Human kidney tissue lysate, 30 µg/Lane Lane 4: Mouse liver tissue lysate, 30 µg/Lane Lane 5: Mouse kidney tissue lysate, 30 µg/Lane Lane 6: Rat liver tissue lysate, 30 µg/Lane Lane 7: Rat kidney tissue lysate, 30 µg/Lane
Exposure time: 6 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: EM1701-87, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 46.5 kDa Observed band size: 47 kDa
☑ Knockdown (KD)
Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (EM1701-87) at 1/2,000 dilution.
Lane 1: A549-WT cell lysate Lane 2: A549-KD ASS1 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 47 kDa Observed band size: 45 kDa
Exposure time: 40 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 (EM1701-87) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining ASS1 (green) in MCF-7 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC staining ASS1 (green) in SiHa cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC staining ASS1 (green) in HepG2 cells. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-ASS1 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-ASS1 antibody. Counter stained with hematoxylin.
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