Name: Anti-CX3CR1 (extracellular) Antibody
Brand: Alomone Labs
SKU: ACR-058

Overview

Cat #: ACR-058

Alternative Name Fractalkine Receptor, CX3C chemokine receptor 1, C-X3-C CKR-1, C-X3-C motif chemokine receptor 1, GPR13, CMKBRL1, GPRV28, V28, G-protein coupled receptor 13, CCRL1, Beta chemokine receptor-like 1

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: m, r

Immunogen

Peptide (C)ELDLENFEYDDSAE, corresponding to amino acid residues 7- 20 of mouse CX3CR1 (Accession Q9Z0D9). Extracellular, N-terminus.

Accession (Uniprot) Number Q9Z0D9

Gene ID 13051

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Rat – identical.

RRID AB_2876846

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4.

Isotype Rabbit IgG

Specificity Not recommended for use with human samples.

Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.

Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.

Reconstitution 0.2 ml double distilled water (DDW).

Antibody concentration after reconstitution 0.8 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).

Standard quality control of each lot Western blot analysis.

Applications: ifc, ih, lci, wb

May also work in: ic*, ip*

Western blot

Western blot analysis of rat brain lysate (lanes 1 and 4), mouse brain membranes (lanes 2 and 5) and rat spleen lysate (lanes 3 and 6):
1-3. Anti-CX3CR1 (extracellular) Antibody (#ACR-058), (1:200).
4-6. Anti-CX3CR1 (extracellular) Antibody, preincubated with CX3CR1 (extracellular) Blocking Peptide (#BLP-CR058).
Western blot analysis of mouse BV-2 microglia cell line lysate (lanes 1 and 3) and mouse J774 macrophage cell line lysate (lanes 2 and 4):
1, 2. Anti-CX3CR1 (extracellular) Antibody (#ACR-058), (1:200).
3, 4. Anti-CX3CR1 (extracellular) Antibody, preincubated with CX3CR1 (extracellular) Blocking Peptide (#BLP-CR058).

Immunohistochemistry

Expression of CX3CR1 in rat hippocampus
Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-CX3CR1 (extracellular) Antibody (#ACR-058), (1:300), followed by goat anti-rabbit-AlexaFluor-488. A. CX3CR1 staining (green) in the hippocampal CA1 region is detected in microglial profiles in the stratum oriens. Some profiles include the entire microglial outline including soma (horizontal arrows) while other outlines are processes (vertical arrows). B. Pre-incubation of the antibody with CX3CR1 (extracellular) Blocking Peptide (#BLP-CR058), suppresses staining. Cell nuclei are stained with DAPI (blue). P = pyramidal layer, SR = stratum radiatum, SO = stratum oriens.
Multiplex staining of CX3CR1 and CD11b in rat hippocampus.
Immunohistochemical staining of perfusion-fixed frozen rat brain sections Anti-CX3CR1 (extracellular) Antibody (#ACR-058), (1:300), followed by goat anti-rabbit-AlexaFluor-488 and mouse Anti -CD11b, (1:500), followed by goat anti-mouse-biotin and streptavidin-Cy3. A. CXCR1 immunoreactivity (green) appears in microglial processes (up pointing arrows) and in full microglial outline (down pointing arrows). B. CD11b immunoreactivity (red) appears in microglial profiles. C. Merge of the two images demonstrates ubiquitous co-localization. Cell nuclei are stained with DAPI (blue). P = pyramidal layer, SR = stratum radiatum.

Indirect flow cytometry

Cell surface detection of CX3CR1 in live intact mouse BV-2 microglia cells:
^___ Cells.
^___ Cells + goat-anti-rabbit-FITC.
^___ Cells + Anti-CX3CR1 (extracellular) Antibody (#ACR-058), (2.5 µg) + goat-anti-rabbit-FITC.

References

Bronger, H. et al. (2019) Cancer Metastasis Rev. 38, 417.
Hamon, P. et al. (2017) Blood 129, 1296.
Luo, P. et al. (2019) Brain. Res. Bull. 146, 12.
Lee, M. et al. (2018) Immune Netw. 18, e5.
Asai, H. et al. (2015) Nat. Neurosci. 18, 1584.

Scientific background

CX₃C-chemokine receptor 1 (CX₃CR1) is a 368 amino acid (aa) protein that contains seven-transmembrane domains and belongs to the family of G protein coupled receptors¹.

CX₃CR1 has a central role in adhesion and chemotaxis through binding to its ligand, fractalkine CX₃C chemokine ligand 1, (CX₃CL1). The major role of CX₃CR1 in immune cells is to recognize and enter inflamed tissue according to CX₃CL1 gradient and initiate the innate immune system². The receptor-ligand axis also mediates the crosstalk between glial cells and neurons by direct or indirect ways in the central nervous system (CNS)³. Expression and interaction of CX₃CR1 with CX₃CL1 is related to many diseases among which are asthma, cancer and gut infections (reviewed in 4).

Both CX₃CL1 and CX₃CR1 are expressed throughout the body. However, their expression is highly cell type-specific depending on organs and tissues. In the brain, CX₃CL1/CX₃CR1 signaling can modulate the production of cytokines by microglia cells. It has been reported that CX₃CL1/CX₃CR1 signaling is associated with Alzheimer's Disease⁵. In the liver, CX₃CR1 is expressed in monocytes, CD8⁺ T cells, and natural killer (NK) cells. Besides immune cells, it is highly expressed at inflammatory sites⁴.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Lyophilized Powder

Anti-CX3CR1 (extracellular) Antibody (#ACR-058) is a highly specific antibody directed against an epitope of the mouse receptor. The antibody can be used in western blot, immunohistochemistry, and indirect flow cytometry applications. It has been designed to recognize CX3CR1 from mouse and rat samples. The antibody will not recognize the protein from human samples.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 29/10/2024

Applications

Citations

Specifications

Scientific Background

Specific Control Product

CX3CR1 (extracellular) Blocking Peptide (#BLP-CR058) is the original antigen used for immunization during Anti-CX3CR1 (extracellular) Antibody (#ACR-058) generation. The blocking peptide binds and ‘blocks’ Anti-CX3CR1 (extracellular) primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.

CX3CR1 (extracellular) Blocking Peptide (#BLP-CR058)

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Anti-CX3CR1 (extracellular) Antibody