CX3CR1 Antibody

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ProSci
Product Code: PSI-2093
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2093-0.02mg0.02mg£150.00
Quantity:
PSI-2093-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Flow Cytometry
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation in Human Cells </strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 KD Validation in 293 Cells</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 293 cells transfected with control siRNAs. Lane 2: 293 cells transfected with CX3CR1 siRNAs.
3 / 15
<strong>Figure 3 Western Blot Validation in THP1 Cells </strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 0.2 μg/mL Lane 1: 0.5 μg/mL Lane 1: 1 μg/mL
4 / 15
<strong>Figure 4 Western Blot Validation in Human Spleen Lysates</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (1 μg/mL) in the absence (lane 1) or presence of blocking peptide (lane 2), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
5 / 15
<strong>Figure 5 Western Blot Validation in Rat Spleen Tissue</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093, (A; 1 μg/mL, B; 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
6 / 15
<strong>Figure 6 Flow Cytometry Validation of CX3CR1 in THP-1 Cells</strong><br> Overlay histogram showing THP-1 cells stained with 2093 (red line, 1μg/1x10<sup>6</sup> cells). 1 h incubation at 4˚C in 2% FBS/PBS. Followed by secondary antibody 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 1 h 4˚C. <br> <br> Isotype control antibody (Green line) was rabbit IgG1 (1μg/1x10<sup>6</sup> cells) used under the same conditions. Acquisition of >10,000 events was performed.
7 / 15
<strong>Figure 7 Immunofluorescence Validation of CX3CR1 In K562 Cells </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling CX3CR1 with 2093 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue). Image showing membrane staining on K562 cells.
8 / 15
<strong>Figure 8 Immunofluorescence Validation of CX3CR1 in Human Heart </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human heart tissue labeling CX3CR1 with 2093 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
9 / 15
<strong>Figure 9 Immunofluorescence Validation of CX3CR1 in Mouse Heart Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse heart tissue labeling CX3CR1 with 2093 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
10 / 15
<strong>Figure 10 Immunohistochemistry Validation of CX3CR1 in Rat Heart Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-CX3CR1 antibody (2093) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
11 / 15
<strong>Figure 11 Immunofluorescence Validation of CX3CR1 in Rat Heart Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed rat heart tissue labeling CX3CR1 with 2093 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
12 / 15
<strong>Figure 12 Immunofluorescence Validation of CX3CR1 in Human Atherosclerosis (Lucas et al., 2003)</strong><br> CX3CR1-positive cells in coronary artery sections was detected by anti-CX3CR1 antibodies by double staining with an anti?smooth muscle actin antibody (yellow).
13 / 15
<strong>Figure 13 Regulated Expression Validation of CX3CR1 in Mouse Bone Marrow-derived NK Cells (Barlic et al., 2003)</strong><br> Western blot analysis of CX3CR1 expression detected by anti-CX3CR1 antibodies was induced by IL-2 but was inhibited by IL-15. Mouse skeletal muscle was for negative control and mouse brain was for positive control.
14 / 15
<strong>Figure 14 Flow Cytometry Validation of CX3CR1 in Peripheral Blood Mononuclear Cells (PBMCs) of Patients (Yajima et al., 2005)</strong><br> Flow cytometry analysis of CX3CR1 expression detected by anti-CX3CR1 antibodies. Protein expression was more pronounced on CD4<sup>+</sup> and CD8<sup>+</sup> T cells from patients with untreated active systemic lupus erythematosus (SLE) as compared to those with rheumatoid arthritis (RA) or healthy controls.
15 / 15
<strong>Figure 15 CX3CR1 Deficiency Validation in Transgenic Mice (Kumar et al., 2013)</strong><br> Transgenic CX3CR1 gfp (C57BL6/J background) mice, in which either one (CX3CR1 gfp/+ ) or both (CX3CR1 gfp/gfp ) copies of the CX3CR1 gene were interrupted by fluorescent protein (GFP). More CX3CR1 positive cells (green) are found in vascular wall of CX3CR1 gfp/+ mice, but not in CX3CR1 gfp/gfp mice, where cells remained in perivascular region due to leaky microvessels. CX3CR1 expression was detected by anti-CX3CR1 antibodies.
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<strong>Figure 1 Western Blot Validation in Human Cells </strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 KD Validation in 293 Cells</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 293 cells transfected with control siRNAs. Lane 2: 293 cells transfected with CX3CR1 siRNAs.
<strong>Figure 3 Western Blot Validation in THP1 Cells </strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 0.2 μg/mL Lane 1: 0.5 μg/mL Lane 1: 1 μg/mL
<strong>Figure 4 Western Blot Validation in Human Spleen Lysates</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093 (1 μg/mL) in the absence (lane 1) or presence of blocking peptide (lane 2), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Western Blot Validation in Rat Spleen Tissue</strong><br> Loading: 15 μg of lysates per lane. Antibodies: CX3CR1 2093, (A; 1 μg/mL, B; 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 6 Flow Cytometry Validation of CX3CR1 in THP-1 Cells</strong><br> Overlay histogram showing THP-1 cells stained with 2093 (red line, 1μg/1x10<sup>6</sup> cells). 1 h incubation at 4˚C in 2% FBS/PBS. Followed by secondary antibody 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 1 h 4˚C. <br> <br> Isotype control antibody (Green line) was rabbit IgG1 (1μg/1x10<sup>6</sup> cells) used under the same conditions. Acquisition of >10,000 events was performed.
<strong>Figure 7 Immunofluorescence Validation of CX3CR1 In K562 Cells </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling CX3CR1 with 2093 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue). Image showing membrane staining on K562 cells.
<strong>Figure 8 Immunofluorescence Validation of CX3CR1 in Human Heart </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human heart tissue labeling CX3CR1 with 2093 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 9 Immunofluorescence Validation of CX3CR1 in Mouse Heart Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse heart tissue labeling CX3CR1 with 2093 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 10 Immunohistochemistry Validation of CX3CR1 in Rat Heart Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-CX3CR1 antibody (2093) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 11 Immunofluorescence Validation of CX3CR1 in Rat Heart Tissue</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed rat heart tissue labeling CX3CR1 with 2093 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 12 Immunofluorescence Validation of CX3CR1 in Human Atherosclerosis (Lucas et al., 2003)</strong><br> CX3CR1-positive cells in coronary artery sections was detected by anti-CX3CR1 antibodies by double staining with an anti?smooth muscle actin antibody (yellow).
<strong>Figure 13 Regulated Expression Validation of CX3CR1 in Mouse Bone Marrow-derived NK Cells (Barlic et al., 2003)</strong><br> Western blot analysis of CX3CR1 expression detected by anti-CX3CR1 antibodies was induced by IL-2 but was inhibited by IL-15. Mouse skeletal muscle was for negative control and mouse brain was for positive control.
<strong>Figure 14 Flow Cytometry Validation of CX3CR1 in Peripheral Blood Mononuclear Cells (PBMCs) of Patients (Yajima et al., 2005)</strong><br> Flow cytometry analysis of CX3CR1 expression detected by anti-CX3CR1 antibodies. Protein expression was more pronounced on CD4<sup>+</sup> and CD8<sup>+</sup> T cells from patients with untreated active systemic lupus erythematosus (SLE) as compared to those with rheumatoid arthritis (RA) or healthy controls.
<strong>Figure 15 CX3CR1 Deficiency Validation in Transgenic Mice (Kumar et al., 2013)</strong><br> Transgenic CX3CR1 gfp (C57BL6/J background) mice, in which either one (CX3CR1 gfp/+ ) or both (CX3CR1 gfp/gfp ) copies of the CX3CR1 gene were interrupted by fluorescent protein (GFP). More CX3CR1 positive cells (green) are found in vascular wall of CX3CR1 gfp/+ mice, but not in CX3CR1 gfp/gfp mice, where cells remained in perivascular region due to leaky microvessels. CX3CR1 expression was detected by anti-CX3CR1 antibodies.

Documents

Further Information

Additional Names:
CX3CR1 Antibody: V28, CCRL1, GPR13, CMKDR1, GPRV28, CMKBRL1, CX3C chemokine receptor 1, Beta chemokine receptor-like 1, C-X3-C CKR-1
Application Note:
WB: 1 -2 μg/mL; IF: 10-20 μg/mL, IHC-P: 2 μg/mL: Flow: 0.1 μg/mL .

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in rat samples; Immunofluorescence in human, mouse and rat samples; Flow cytometry in human samples. All other applications and species not yet tested.
Background:
CX3CR1 Antibody: CX3CR1 is one of the chemokine receptors that are required as coreceptors for HIV infection. The genes encoding human, murine, and rat CX3CR1 were cloned and designated V28 and CMKBRL1, CX3CR1, and RBS11, respectively. The encoded seven transmembrane protein was recently identified as the receptor for a novel transmembrane molecule, fractalkine, and renamed CX3CR1. Recently, CX3CR1 was found to serve as a coreceptor for HIV-1 and HIV-2 envelope fusion and virus infection, which can be inhibited by fractokine. CX3CR1 mediates leukocyte migration and adhesion. CX3CR1 is expressed in a variety of human tissues and cell lines.
Background References:
  • Raport et al. Gene 1995;163:295-9.
  • Combadiere et al. DNA Cell Biol 1995;14:673-80.
  • Combadiere et al. Biochem Biophys Res Commun 1998;253:728-32.
  • Harrison et al. Neurosci Lett 1994;169:85-9.
Buffer:
CX3CR1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Bovine: (82%)
Immunogen:
Anti-CX3CR1 antibody (2093) was raised against a peptide corresponding to 20 amino acids near the amino terminus of mature human CX3CR1.

The immunogen is located within the first 50 amino acids of CX3CR1.
ISOFORMS:
Human CX3CR1 has 3 isoforms, including isoform 1 (355aa, 40kD), isoform 2 (387aa, 44kD), isoform 3 (362aa, 41kD). Mouse CX3CR1 has one isoform (354aa, 40kD) and rat CX3CR1 also has one isoform (354aa, 40kD). 2093 can detect all isoforms of human, mouse and rat.
NCBI Gene ID #:
1524
NCBI Official Name:
chemokine (C-X3-C motif) receptor 1
NCBI Official Symbol:
CX3CR1
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 40-44 kD

Observed: 50 kD
Protein Accession #:
NP_001328
Protein GI Number:
4503171
Purification:
CX3CR1 Antibody is affinity chromatography purified via peptide column.
Research Area:
Chemokines & Cytokines,Cancer,Infectious Disease
Swissprot #:
P49238
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

KD Validation (Figure 2): Anti-CX3CR1 antibody (2093) specificity was further verified by CX3CR1 specific knockdown. CX3CR1 signal in 293 cells transfected with CX3CR1 siRNAs was disrupted in comparison with that in cells transfected with control siRNAs.

Regulated expression validation (Figure 13): CX3CR1 expression detected by anit-CX3CR1 antibodies (2093) was up-regulated by IL-2, which was inhibited by IL-15.

Deficiency Validation (Figure 15): CX3CR1+ cells detected by anti-CX3CR1 antibodies is localized in vascular wall of CX3CR1+/gfp mice, but not in CX3CR1gfp/gfp mice, where cells remained in perivascular region.

References

  1. Lucas et al. Smooth muscle cells in human atherosclerotic plaques express the fractalkine receptor CX3CR1 and undergo chemotaxis to the CX3C chemokine fractalkine (CX3CL1). Circulation. 2003;108(20):2498-504.PMID: 14581400
  2. Barlic et al. IL-15 and IL-2 oppositely regulate expression of the chemokine receptor CX3CR1. Blood. 2003;102(10):3494-503. PMID: 12881312
  3. Yajima et al. Elevated levels of soluble fractalkine in active systemic lupus erythematosus: potential involvement in neuropsychiatric manifestations. Arthritis Rheum. 2005;52(6):1670-5. PMID: 15934075
  4. Kumar et al. Role of CX3CR1 receptor in monocyte/macrophage driven neovascularization. PLoS One. 2013;8(2):e57230. PMID: 23437346
  5. Beutner et al. Engineered stem cell-derived microglia as therapeutic vehicle for experimental autoimmune encephalomyelitis. Gene Ther. 2013;20(8):797-806. PMID: 23324824
  6. Wang et al. TGF-?/Smad3 signalling regulates the transition of bone marrow-derived macrophages into myofibroblasts during tissue fibrosis. Oncotarget. 2016;7(8):8809-22. PMID: 26684242
  7. Kang et al. Intestinal epithelial cell-derived semaphorin 7A negatively regulates development of colitis via ?v?1 integrin. J Immunol. 2012;188(3):1108-16. PMID: 22198947
  8. Umemura et al. Tumor-infiltrating myeloid-derived suppressor cells are pleiotropic-inflamed monocytes/macrophages that bear M1- and M2-type characteristics. J Leukoc Biol. 2008;83(5):1136-44. PMID: 18285406
  9. You et al. Fractalkine, a CX3C chemokine, as a mediator of ocular angiogenesis. Invest Ophthalmol Vis Sci. 2007;48(11):5290-8. PMID: 17962485
  10. Matsunawa et al. Elevated serum levels of soluble CX3CL1 in patients with microscopic polyangiitis. Clin Exp Rheumatol. 2009;27(1):72-8. PMID: 19327232
  11. Sato et al. Involvement of CX3CL1/CX3CR1 axis in etanercept therapy for patients with active rheumatoid arthritis. Open Access Rheumatol. 2011;3:1-7. PMID: 27789999
  12. Clara Beutner. Microglia derived from embryonic stem cells and its application in CNS diseases. PhD Thesis. University of Bonn, Germany. 2012.PMID:
  13. Kristin Roy. Establishment of microglial precursors derived from human induced pluripotent stem cells to model SOD1-mediated amyotrophic lateral sclerosis. PhD Thesis. University of Bonn, Germany. 2012.PMID:
  14. Isabella Napoli. Establishment of Embryonic Stem Cell Derived Microglial Precursors and Application in an Animal Model of Alzheimer?s Disease. PhD Thesis. University of Bonn, Germany. 2008.PMID:
  15. Kalaimathi Govindarajan. Role of KLF4 in regulation of myocardin induced SMC differentiation in human smooth muscle stem progenitor cells (hSMSPC). PhD Thesis. University of College Cork, Ireland. 2013.PMID:

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