Mouse CCL12/MCP-5 Antibody (63205)

Chemotaxis Induced by CCL12/MCP‑5 and Neutral-ization by Mouse CCL12/ MCP‑5 Antibody.

Recombinant Mouse CCL12/MCP-5 (Catalog # 428-P5) chemoattracts the BaF3 mouse pro-B cell line transfected with human CCR2A 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 CCL12/ MCP-5 (10 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse CCL12/MCP-5 Monoclonal Antibody (Catalog # MAB428). The ND₅₀ is typically 0.5-2.5 µg/mL. Maximum neutralization observed was around 80%.

Mouse CCL12 / MCP-5 ELISA Standard Curve

Recombinant Mouse CCL12/MCP‑5 (Catalog # 428-P5) was serially diluted and captured by Rat Anti-Mouse CCL12/MCP‑5 Monoclonal Antibody (Catalog # MAB428) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse CCL12/MCP‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF428) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Detection of CCL12/MCP-5 by Flow Cytometry

Integrated analysis of chemokine/cytokine protein after 9-Gy myeloablative radiation, HSCs, and adoptive cellular therapy and in vitro migration assays and single-cell RNA sequencing analysis in KR158B glioma. (A) Cytokine and chemokine profiling of glioma protein lysates in the tumor secretome in adoptive cellular therapy–treated gliomas and untreated control. Statistical comparisons were made using the 1-way ANOVA test. (B) CCL12 protein levels in glioma lysates were quantified by ELISA. Statistical analysis was made using the unpaired Student t test. (C) Transcriptomic analysis of chemokine ligands and receptors using nanoString digital spatial profiling. Statistical analysis was performed using the 2-way ANOVA test. (D) In vitro trans-well migration assay of MDSCs migrating in response to glioma conditioned media from excised glioma tissue combined with neutralizing antibodies against CCL2 and/or CCL12. Statistical analysis was performed using the 1-way ANOVA test. (E) ELISA CCL12 protein quantification using immune cell–specific isolates from excised glioma tissue. Statistical analysis was performed using the 1-way ANOVA test. (F) Immune cell subset deconvolution from single-cell RNA sequencing analysis from excised glioma tissue. (G) mRNA expression of CCL12 overlayed on immune cell subset deconvolution using single-cell RNA sequencing analysis from excised glioma tissue. (H) In vitro trans-well migration assay of MDSCs migration in response to immune cell-specific isolates combined with either IgG or CCL12 neutralizing antibody. Statistical comparisons conducted using the 2-way ANOVA test. *P ≤.05. **P ≤.01. ***P ≤.001. ****P ≤.0001; error bars: mean ± standard deviation. Abbreviations: ANOVA, analysis of variance; MDSCs, myeloid-derived suppressor cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41908149), licensed under a CC-BY license. Not internally tested by R&D Systems.