Recombinant Human ARF1 protein(Met1-Lys181), GST-tagged

• Cat.No.: ARF1-9799H
• Product Overview: Recombinant Human ARF1 (P84077) (Met1-Lys181) was expressed in E. coli with a GST tag at the N-terminus.

Specification

• Species: Human
• Source: E.coli
• Tag: GST
• Protein Length: Met1-Lys181
• Form: Lyophilized from sterile PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human ARF1 consists of 407 amino acids and predicts a molecular mass of 47.00 kDa.
• Endotoxin: < 1.0 EU per μg protein as determined by the LAL method.
• Purity: ≥ 95 % as determined by SDS-PAGE. ≥ 90 % as determined by SEC-HPLC.
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: ARF1 ADP-ribosylation factor 1 [ Homo sapiens ]
• Official Symbol: ARF1
• Synonyms: ARF1; ADP-ribosylation factor 1
• Gene ID: 375
• mRNA Refseq: NM_001024226
• Protein Refseq: NP_001019397
• MIM: 103180
• UniProt ID: P84077

Case Study

Case 1: Enkler L, et al. Nat Cell Biol. 2023

Fatty acid β-oxidation is key for energy making when nutrients are low. In yeast, this starts in the peroxisome, then the products head to mitochondria to fuel the tricarboxylic acid cycle. We discovered that a hyperactive mutant of the small GTPase Arf1 lowers the expression of fatty acid transporters and the key enzyme for β-oxidation. This causes fatty acids to pile up in lipid droplets, resulting in fragmented mitochondria and reduced ATP production. The same mitochondrial issues appeared when fatty acids were depleted, whether by genetics or drugs. While β-oxidation happens in both mitochondria and peroxisomes in mammals, Arf1's impact on fatty acid metabolism is a shared trait.

Fig1. Single timepoint images of movies done with strains expressing yArf1–GFP.

Fig2. Active yArf1 pull-down and detection experiments done with strains expressing yArf1 and yArf1-11 fused to GFP.

Case 2: Kotani Y, et al. Sci Rep. 2022

Mast cells are key in allergies, releasing inflammatory substances when activated via the IgE receptor (FcεRI), a process involving vesicle movement. The GTPase Arf1, which manages membrane traffic and cell structure, likely aids this activation. Blocking Arf1 with drugs can curb cytokine release and degranulation in mast cells. Yet, its exact role is fuzzy. Through genetic studies, we've shown Arf1 is vital for mTORC1 activation when IL-3 is present and aids mast cell growth. Surprisingly, lacking Arf1 doesn't much affect reactions like degranulation or cytokine output triggered by FcεRI. This unveils Arf1's unexpected part in mast cell multiplication and suggests its potential as a target in disorders where mast cells proliferate excessively.

Fig1. Either control (ctrl) or Arf1-KO BMMCs at 5-week after culture were stimulated with the indicated concentration of IL-3.

Fig2. Control (ctrl) or Arf1-KO BMMCs sensitized with anti-DNP IgE were stimulated with the indicated concentrations of DNP-BSA.

Application

Recombinant Human ARF1 (rhARF1) is quite handy in cell and molecular biology studies. Part of the ADP-ribosylation factor family, this small GTPase helps manage protein transport in the Golgi apparatus. Researchers use rhARF1 to dig into its vital roles in Golgi vesicle formation and shedding, along with how it controls protein movement and membrane organization in cells. Moreover, ARF1 engages in RNA transcription and protein translation by enhancing the connection between RNA polymerase and DNA, supporting RNA synthesis. Hence, rhARF1 is valuable for delving into cell signaling, protein production, and organelle functions.

In industrial realms, rhARF1 finds its niche in biopharmaceuticals and drug creation. It's a potential drug target for developing inhibitors of ARF1 enzyme activity that might treat diseases like cancer. RhARF1 is also used to create antibodies for scientific studies, including immunohistochemistry, Western Blot, immunocytochemistry, ELISA, and flow cytometry. These antibodies help detect and study ARF1 expression and functionality in samples from different species. Overall, its applications aid in grasping ARF1's biological roles and pave the way for new therapeutic developments.

Fig1. Regulation of actin assembly by ARF1 during vesicle formation at the Golgi. (Crislyn D'Souza-Schorey, 2006)