Recombinant Human ARF3 protein(Met1-Lys181), His-tagged

• Cat.No.: ARF3-237H
• Product Overview: Recombinant Human ARF3 (P61204) (Met1-Lys181) was expressed in E. coli with a polyhistidine tag at the N-terminus.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Protein Length: 1-181 a.a.
• Form: Lyophilized from sterile 50mM Tris, 10% glycerol, pH 8.0. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human ARF3 consists of 196 amino acids and predicts a molecular mass of 22.4 KDa. It migrates as an approximately 23 KDa band in SDS-PAGE under reducing conditions.
• Purity: > 85 % as determined by SDS-PAGE
• 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: ARF3 ADP-ribosylation factor 3 [ Homo sapiens ]
• Official Symbol: ARF3
• Synonyms: ARF3; ADP-ribosylation factor 3; small GTP binding protein
• Gene ID: 377
• mRNA Refseq: NM_001659
• Protein Refseq: NP_001650
• MIM: 103190
• UniProt ID: P61204

Case Study

Case 1: Sandilands E, et al. J Cell Biol. 2023

ARF GTPases are key players in managing membrane trafficking and helping in vesicle creation. Understanding their role is tricky due to their interactions with various proteins like GEFs, GAPs, and others. In a study on 3D prostate cancer cell behavior, we checked how these components influence the way cancer cells invade. We found that ARF3 GTPase plays a crucial part in determining how invasion occurs, either through leader-led chains or as a group sheet movement. ARF3's control over invasion is linked to its influence on the turnover of N-cadherin. In live animals, ARF3 levels can adjust metastasis tendencies from prostate tumors. Tracking ARF3 and N-cadherin levels can help identify prostate cancer cases with aggressive outcomes. This research highlights a special function of ARF3 in organizing cell behavior during invasion and cancer spread.

Fig1. Western blot of androgen receptor (AR) proficient or deficient prostate cell lines for ARF1, ARF3, ARF4, ARF5, and ARF6.

Fig2. PC3 cells expressing mNG, ARF1-mNG, or ARF3-mNG and Scr shRNA were classified into Round, Spindle, and Spread.

Case 2: Li F, et al. FEBS Lett. 2002

To figure out which genes get influenced by heregulin-beta1 (HRG) in breast cell regulation, we checked out the mRNA of MCF-7 cells. We discovered that HRG ramps up the levels of a clone matching human ADP-ribosylation factor 3 (ARF3), which is a protein involved in vesicle movement and cell transport. When cells were treated with HRG, both ARF3 mRNA and protein levels went up. Interestingly, ARF3 quickly moved to cell membranes and then into the nucleus, lining up with acetylated histone H3 in certain spots. Moreover, ARF3 levels varied in the mammary gland, peaking in virgin and post-weaning stages. These insights unveil a new role for ARF3 in how HRG operates in mammary epithelial cells, hinting at its involvement in cellular processes influenced by HRG.

Fig1. HRG induces ARF3 mRNA expression in the absence of de novo protein synthesis.

Fig2. ARF3 protein expression in breast cancer cells as determined by Western blot.

Application

Recombinant Human ARF3 (rhARF3), primarily used in research, is all about diving deep into how cells shuttle stuff around and send signals. You can think of ARF3 as a member of the small GTPase family, playing a crucial role in moving things around cell membranes, creating vesicles, and engaging in endocytosis. Researchers are keen on using rhARF3 to figure out how it gets busy when it binds to GTP—basically kickstarting vesicle formation and endocytosis. Plus, it's super important for transferring materials from the endoplasmic reticulum to the Golgi apparatus. ARF3 also has a hand in the endocytosis of cell surface receptors and signaling pathways. Unraveling how ARF3 ticks in different cell types and seeing what changes under disease conditions are pretty big deals.

When we shift gears to the industrial production side of things, rhARF3 finds its place in biopharmaceuticals and drug development. Since any hiccups in ARF3's function can throw cellular transport into chaos and link up with certain metabolic disorders, targeting rhARF3 could pave the way for drugs that inhibit its enzyme activity. Imagine having meds that might help treat certain cancers! Moreover, rhARF3 makes waves in producing antibodies for research. These antibodies are handy for a bunch of scientific experiments like immunohistochemistry, Western Blot, immunocytochemistry, ELISA, and flow cytometry, helping in detecting and studying ARF3 across different samples. All these applications not only boost our grasp of ARF3's biological roles but also open new doors for therapeutic breakthroughs.

Fig1. Schematic illustration of the signaling mechanisms by which both ARF3 and ARF6 regulate CpG ODN/TLR9-mediated signaling. (Jing-Yiing Wu, 2015)