Mouse CX3CL1/Fractalkine Chemokine Domain Antibody

Chemotaxis Induced by CX3CL1/Fractalkine and Neutralization by Mouse CX3CL1/Fractalkine Antibody. Recombinant Mouse CX3CL1/Fractalkine (Catalog # 571-MF) chemoattracts the BaF3 mouse pro-B cell line transfected with human CX3CR1 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Mouse CX3CL1/Fractalkine (30 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse CX3CL1/Fractalkine Chemokine Domain Monoclonal Antibody (Catalog # MAB571). The ND₅₀ is typically 0.75-3.5 µg/mL.

Detection of Mouse CX3CL1/Fractalkine by Immunocytochemistry/Immunofluorescence CX3CL1 expression in the brain during EAE progression. (A) Brain CX3CL1 expression of wild type mice with EAE as determined by quantitative real-time PCR. Gene levels were normalized to GAPDH levels and are displayed as levels relative to naïve mice. Error bars represent the s.e.m. (n = 5 mice per group). (B-M) Brains from naïve wild type mice were harvested and frozen for immunostaining. CX3CL1 expression (red) and DAPI nuclei staining (blue) in (B, F, J) naïve and (C, G, K) Day 10, (D, H, L) Day 14, and (E, I, M) Day 21 post-EAE induced wild type mice. CX3CL1 expression is displayed at the (B-E) choroid plexus, (F-I) in and near the hippocampus, and (J-M) cerebellum. White scale bars represent 50 μm. (N) CX3CL1 expression (relative to non-treated cells) in the CPLacZ-2 mouse choroid plexus cell line after 2 hour treatment with varying concentrations of the A2A adenosine receptor specific adenosine receptor agonist CGS21680. Error bars represent the s.e.m. (O) Lymphocyte migration across a transwell choroid plexus barrier following pretreatment with vehicle treatment alone, CGS21680, or CGS21680 and anti-CX3CL1. Total migration was normalized to the vehicle control (set to 100%). Error bars represent the s.e.m. These results are representative of two separate experiments (n ≤ 3). Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CX3CL1/Fractalkine by Immunohistochemistry-Frozen CX3CL1 antibody mediated blockade protects mice against EAE and its associated lymphocyte infiltration. Wild type mice were induced to develop EAE and starting at day 8 post induction given daily anti-CX3CL1 antibody or an isotype control treatments (i.p.). (A) EAE disease profile. Error bars represent the s.e.m. (n = 4 mice/group). Significant differences are indicated as determined by two-way ANOVA. EAE scoring data is representative of 2 separate experiments. (B) CD45, CD11b, and F480 stained brain (hippocampal and cerebellum areas) and spinal cord sections from day 28 post-EAE induced mice treated with either anti-CX3CL1 or control antibody. Positively stained cells (red) are shown against a hematoxylin counterstain (blue). Black scale bars represent 50 μm. (C) CD4 and (D) CD8 positive mean cells counts per field at 10x magnification from brain and spinal cord stained frozen brain sections from day 28 post-EAE induced mice treated with either anti-CX3CL1 or control antibody. Error bars represent the standard error of the mean (n ≤ 11). Significant differences (P < 0.05, *) are indicated as determined by the Student’s t-test. Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CX3CL1/Fractalkine by Immunocytochemistry/Immunofluorescence CX3CL1 expression in the brain during EAE progression. (A) Brain CX3CL1 expression of wild type mice with EAE as determined by quantitative real-time PCR. Gene levels were normalized to GAPDH levels and are displayed as levels relative to naïve mice. Error bars represent the s.e.m. (n = 5 mice per group). (B-M) Brains from naïve wild type mice were harvested and frozen for immunostaining. CX3CL1 expression (red) and DAPI nuclei staining (blue) in (B, F, J) naïve and (C, G, K) Day 10, (D, H, L) Day 14, and (E, I, M) Day 21 post-EAE induced wild type mice. CX3CL1 expression is displayed at the (B-E) choroid plexus, (F-I) in and near the hippocampus, and (J-M) cerebellum. White scale bars represent 50 μm. (N) CX3CL1 expression (relative to non-treated cells) in the CPLacZ-2 mouse choroid plexus cell line after 2 hour treatment with varying concentrations of the A2A adenosine receptor specific adenosine receptor agonist CGS21680. Error bars represent the s.e.m. (O) Lymphocyte migration across a transwell choroid plexus barrier following pretreatment with vehicle treatment alone, CGS21680, or CGS21680 and anti-CX3CL1. Total migration was normalized to the vehicle control (set to 100%). Error bars represent the s.e.m. These results are representative of two separate experiments (n ≤ 3). Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CX3CL1/Fractalkine by Immunocytochemistry/Immunofluorescence CX3CL1 expression in the brain during EAE progression. (A) Brain CX3CL1 expression of wild type mice with EAE as determined by quantitative real-time PCR. Gene levels were normalized to GAPDH levels and are displayed as levels relative to naïve mice. Error bars represent the s.e.m. (n = 5 mice per group). (B-M) Brains from naïve wild type mice were harvested and frozen for immunostaining. CX3CL1 expression (red) and DAPI nuclei staining (blue) in (B, F, J) naïve and (C, G, K) Day 10, (D, H, L) Day 14, and (E, I, M) Day 21 post-EAE induced wild type mice. CX3CL1 expression is displayed at the (B-E) choroid plexus, (F-I) in and near the hippocampus, and (J-M) cerebellum. White scale bars represent 50 μm. (N) CX3CL1 expression (relative to non-treated cells) in the CPLacZ-2 mouse choroid plexus cell line after 2 hour treatment with varying concentrations of the A2A adenosine receptor specific adenosine receptor agonist CGS21680. Error bars represent the s.e.m. (O) Lymphocyte migration across a transwell choroid plexus barrier following pretreatment with vehicle treatment alone, CGS21680, or CGS21680 and anti-CX3CL1. Total migration was normalized to the vehicle control (set to 100%). Error bars represent the s.e.m. These results are representative of two separate experiments (n ≤ 3). Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-193), licensed under a CC-BY license. Not internally tested by R&D Systems.