Recombinant Mouse CX3CL1/Fractalkine (Full Length) Protein

• Reactivity: Mu
• Applications: Bioactivity

Recombinant Mouse CX3CL1/Fractalkine (Full Length) Protein Summary

• Details of Functionality: Measured by its ability to chemoattract freshly isolated human peripheral blood lymphocytes (PBL). The ED₅₀ for this effect is 0.2-0.6 µg/mL. Measured by its ability to chemoattract BaF3 mouse pro‑B cells transfected with mouse CX3CR1. The ED₅₀ for this effect is 0.03-0.12 μg/mL.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse CX3CL1/Fractalkine protein
  Gln25-Arg337 , with a C-terminal 6-His tag
• Accession #: O35188
• N-terminal Sequence: No results obtained: Gln25 predicted
• Protein/Peptide Type: Recombinant Proteins
• Gene: Cx3cl1
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 34 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 90 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse CX3CL1/Fractalkine (Full Length) Protein

• ABCD-3
• C3Xkine
• chemokine (C-X3-C motif) ligand 1
• CX3C membrane-anchored chemokine
• CX3CL1
• CXC3
• CXC3C
• FKN
• Fractalkine
• Neurotactin
• neurotactin)
• NTNsmall inducible cytokine subfamily D (Cys-X3-Cys), member 1 (fractalkine
• NTTSmall-inducible cytokine D1
• SCYD1C-X3-C motif chemokine 1
• small inducible cytokine subfamily D (Cys-X3-Cys), member-1

Background

Fractalkine, designated CX3CL1 and also known as neurotactin, is the only member of the CX3C, or delta, chemokine subfamily (1 ‑ 4). Unlike most other chemokines, CX3CL1 is a type I transmembrane (TM) adhesion protein (1). The mouse CX3CL1 cDNA encodes 395 amino acids (aa), including a signal sequence (aa 1 ‑ 24), a chemokine domain (aa 25 ‑ 100), a mucin stalk region (aa 101 ‑ 336), a transmembrane segment (aa 337 ‑ 357), and a cytoplasmic tail (aa 358 ‑ 395). The chemokine domain contains binding and chemotactic determinants, while the mucin stalk appears to function only as a spacer (4, 5). Mouse CX3CL1 shares 85% and 78% aa sequence identity with rat and human CX3CL1, respectively, within the chemokine domain, but lower sequence identity within other domains. CX3CL1 is up‑regulated by pro‑inflammatory stimuli, especially IFN‑ gamma and TNF‑ alpha , on cell types including macrophages, dendritic cells, endothelium, neurons, smooth muscle and epithelium lining the intestines and other tubules (1, 8, 9). The 40 kDa, 7‑TM non‑glycosylated G‑protein coupled CX3CL1 receptor, CX3CR1, is expressed by cytotoxic effector cells and cytokine producers, including type I helper and cytotoxic T cells, gamma δ T cells, CD16+ NK cells, monocytes and microglia (1, 2). The 95 ‑ 100 kDa TM CX3CL1 can be inducibly cleaved near the TM segment by ADAM10 or ADAM17 to generate a 60 ‑ 80 kDa soluble form (6, 7). TM CX3CL1 functions as an adhesion molecule, while both forms are chemoattractants for target cells expressing CX3CR1 (1, 2). During extravasation, membrane‑bound CX3CL1 traps leukocytes, then is cleaved to allow diapedesis (6). In coronary artery disease, soluble CX3CL1 and CD8+ T cell CX3CR1 are overexpressed and appear to contribute to pathogenesis (1, 10). In the brain, CX3CL1/CX3CR1 interaction protects against microglial neurotoxicity (11). CX3CL1 also contributes to wound healing by recruiting macrophages, and to bone resorption by recruiting and mediating adhesion of osteoclast precursors (12, 13).

1. Stievano, L. et al. (2004) Crit. Rev. Immunol. 24:205.
2. Umehara, H. et al. (2004) Arterioscler. Thromb. Vasc. Biol. 24:34.
3. Rossi, D.L. et al. (1998) Genomics 47:163.
4. Mizoue, L.S. et al. (2001) J. Biol. Chem. 276:33906.
5. Harrison, J.K. et al. (2001) J. Biol. Chem. 276:21632.
6. Hundhausen, C. et al. (2007) J. Immunol. 178:8064.        
7. Tsou, C. et al. (2001) J. Biol. Chem. 276:44622.
8. Tarozzo, G. et al. (2003) J. Neurosci. Res. 73:81.
9. Lucas, A.D. et al. (2001) Am. J. Pathol. 158:855.
10. Damas, J.K. et al. (2005) Arterioscler. Thromb. Vasc. Biol. 25:2567.
11. Cardona, A.E. et al. (2006) Nat. Neurosci. 9:917.
12. Koizumi, K. et al. (2009) J. Immunol. 183:7825.
13. Ishida, Y. et al. (2008) J. Immunol. 180:569.

Publications for CX3CL1/Fractalkine (472-FF)(14)

Publications using 472-FF

• Sokolova, D;Ghansah, SA;Puletti, F;Georgiades, T;De Schepper, S;Zheng, Y;Crowley, G;Wu, L;Rueda-Carrasco, J;Koutsiouroumpa, A;Muckett, P;Freeman, OJ;Khakh, BS;Hong, S; Astrocyte-derived MFG-E8 facilitates microglial synapse elimination in Alzheimer's disease mouse models bioRxiv : the preprint server for biology 2024-09-01 [PMID: 39257734] (Bioassay, Mouse)
• Demetriou, A;Lindqvist, B;Ali, HG;Shamekh, MM;Maioli, S;Inzunza, J;Varshney, M;Nilsson, P;Nalvarte, I; ER? mediates sex-specific protection in the App-NL-G-F mouse model of Alzheimer's disease bioRxiv : the preprint server for biology 2024-07-23 [PMID: 39091856] (Bioassay, Mouse)
• S De Scheppe, JZ Ge, G Crowley, LSS Ferreira, D Garceau, CE Toomey, D Sokolova, J Rueda-Carr, SH Shin, JS Kim, T Childs, T Lashley, JJ Burden, M Sasner, C Sala Frige, S Jung, S Hong Perivascular cells induce microglial phagocytic states and synaptic engulfment via SPP1 in mouse models of Alzheimer&#039;s disease Nature Neuroscience, 2023-02-06;0(0):. 2023-02-06 [PMID: 36747024] (Bioassay, Mouse)
• K Zhou, J Han, H Lund, NR Boggavarap, VM Lauschke, S Goto, H Cheng, Y Wang, A Tachi, C Xie, K Zhu, Y Sun, AM Osman, D Liang, W Han, K Gemzell-Da, C Betsholtz, XM Zhang, C Zhu, M Enge, B Joseph, RA Harris, K Blomgren An overlooked subset of Cx3cr1wt/wt microglia in the Cx3cr1CreER-Eyfp/wt mouse has a repopulation advantage over Cx3cr1CreER-Eyfp/wt microglia following microglial depletion Journal of Neuroinflammation, 2022-01-21;19(1):20. 2022-01-21 [PMID: 35062962] (Bioassay, Mouse)
• EJ Park, PK Myint, MG Appiah, P Worawattan, J Inprasit, O Prajuabjin, ZY Soe, A Gaowa, E Kawamoto, M Shimaoka Ligand-competent fractalkine receptor is expressed on exosomes Biochemistry and Biophysics Reports, 2021-02-02;26(0):100932. 2021-02-02 [PMID: 33553692] (Bioassay, Mouse)
• S Otobe, T Hisamoto, T Miyagaki, S Morimura, H Suga, M Sugaya, S Sato CX3CR1 Deficiency Attenuates DNFB-Induced Contact Hypersensitivity Through Skewed Polarization Towards M2 Phenotype in Macrophages Int J Mol Sci, 2020-10-07;21(19):. 2020-10-07 [PMID: 33036460] (Cell Culture, Mouse)
• M Mizoguchi, Y Ishida, M Nosaka, A Kimura, Y Kuninaka, T Yahata, S Nanjo, S Toujima, S Minami, K Ino, N Mukaida, T Kondo Prevention of lipopolysaccharide-induced preterm labor by the lack of CX3CL1-CX3CR1 interaction in mice PLoS ONE, 2018-11-06;13(11):e0207085. 2018-11-06 [PMID: 30399192] (Bioassay, Mouse)
• Y Zhang, L Zhao, X Wang, W Ma, A Lazere, HH Qian, J Zhang, M Abu-Asab, RN Fariss, JE Roger, WT Wong Repopulating retinal microglia restore endogenous organization and function under CX3CL1-CX3CR1 regulation Sci Adv, 2018-03-21;4(3):eaap8492. 2018-03-21 [PMID: 29750189] (Mouse)
• N Kemmerling, P Wunderlich, S Theil, B Linnartz-G, N Hersch, B Hoffmann, MT Heneka, B de Stroope, H Neumann, J Walter Intramembranous processing by ?-secretase regulates reverse signaling of ephrin-B2 in migration of microglia Glia, 2017-04-03;0(0):. 2017-04-03 [PMID: 28370426] (Bioassay, Mouse)
• Progesterone Attenuates Microglial-Driven Retinal Degeneration and Stimulates Protective Fractalkine-CX3CR1 Signaling PLoS ONE, 2016-11-04;11(11):e0165197. 2016-11-04 [PMID: 27814376] (Bioassay, Mouse)
• McComb JG, Ranganathan M, Liu XH, Pilewski JM, Ray P, Watkins SC, Choi AM, Lee JS CX3CL1 up-regulation is associated with recruitment of CX3CR1+ mononuclear phagocytes and T lymphocytes in the lungs during cigarette smoke-induced emphysema. Am. J. Pathol., 2008-09-04;173(4):949-61. 2008-09-04 [PMID: 18772344] (In Vivo, Mouse)
• Ishida Y, Hayashi T, Goto T, Kimura A, Akimoto S, Mukaida N, Kondo T Essential involvement of CX3CR1-mediated signals in the bactericidal host defense during septic peritonitis. J. Immunol., 2008-09-15;181(6):4208-18. 2008-09-15 [PMID: 18768878] (Bioassay, Mouse)
• Cardona AE, Sasse ME, Liu L, Cardona SM, Mizutani M, Savarin C, Hu T, Ransohoff RM Scavenging roles of chemokine receptors: chemokine receptor deficiency is associated with increased levels of ligand in circulation and tissues. Blood, 2008-03-17;112(2):256-63. 2008-03-17 [PMID: 18347198] (Bioassay, Mouse)
• Lee FH, Haskell C, Charo IF, Boettiger D Receptor-ligand binding in the cell-substrate contact zone: a quantitative analysis using CX3CR1 and CXCR1 chemokine receptors. Biochemistry, 2004-06-08;43(22):7179-86. 2004-06-08 [PMID: 15170355] (Binding Assay, Mouse)

Bioinformatics

• Gene Symbol: Cx3cl1
• Uniprot:
  • O35188()