CX3CR1 Antibody

• Catalog Number: orb1239314
• Category: Antibodies
• Description: CX3CR1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, IF, IHC-P, WB
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-CX3CR1 antibody (orb1239314) 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.
• Concentration: 1 mg/mL
• Dilution range: 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.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 40-44 kDObserved: 50 kD
• Target: CX3CR1
• UniProt ID: P49238
• NCBI: NP_001328
• Storage: CX3CR1 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: CX3CR1 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: CX3CR1 Antibody: V28, CCRL1, GPR13, CMKDR1, GPRV28
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in Human Cells. Loading: 15 µg of lysates per lane. Antibodies: CX3CR1 orb1239314 (0.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

KD Validation in 293 Cells. Loading: 15 µg of lysates per lane. Antibodies: CX3CR1 orb1239314 (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.

Western Blot Validation in THP1 Cells. Loading: 15 µg of lysates per lane. Antibodies: CX3CR1 orb1239314 (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.

Western Blot Validation in Human Spleen Lysates. Loading: 15 µg of lysates per lane. Antibodies: CX3CR1 orb1239314 (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.

Western Blot Validation in Rat Spleen Tissue. Loading: 15 µg of lysates per lane. Antibodies: CX3CR1 orb1239314, (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.

Flow Cytometry Validation of CX3CR1 in THP-1 Cells. Overlay histogram showing THP-1 cells stained with orb1239314 (red line, 1µg/1x10 6 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. Isotype control antibody (Green line) was rabbit IgG1 (1µg/1x10 6 cells) used under the same conditions. Acquisition of > 10000 events was performed.

Immunofluorescence Validation of CX3CR1 In K562 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling CX3CR1 with orb1239314 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.

Immunofluorescence Validation of CX3CR1 in Human Heart. Immunofluorescent analysis of 4% paraformaldehyde-fixed human heart tissue labeling CX3CR1 with orb1239314 at 10 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunofluorescence Validation of CX3CR1 in Mouse Heart Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse heart tissue labeling CX3CR1 with orb1239314 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunohistochemistry Validation of CX3CR1 in Rat Heart Tissue. Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-CX3CR1 antibody (orb1239314) 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.

Immunofluorescence Validation of CX3CR1 in Rat Heart Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed rat heart tissue labeling CX3CR1 with orb1239314 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Immunofluorescence Validation of CX3CR1 in Human Atherosclerosis (Lucas et al., 2003). CX3CR1-positive cells in coronary artery sections was detected by anti-CX3CR1 antibodies by double staining with an anti–smooth muscle actin antibody (yellow).

Regulated Expression Validation of CX3CR1 in Mouse Bone Marrow-derived NK Cells (Barlic et al., 2003). 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.

Flow Cytometry Validation of CX3CR1 in Peripheral Blood Mononuclear Cells (PBMCs) of Patients (Yajima et al., 2005). Flow cytometry analysis of CX3CR1 expression detected by anti-CX3CR1 antibodies. Protein expression was more pronounced on CD4 + and CD8 + T cells from patients with untreated active systemic lupus erythematosus (SLE) as compared to those with rheumatoid arthritis (RA) or healthy controls.

CX3CR1 Deficiency Validation in Transgenic Mice (Kumar et al., 2013). 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.