Recombinant MouseCCL3/MIP‑1 alpha (Catalog #450-MA) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CCR5 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 CCL3/MIP‑1 alpha (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CCL3/MIP‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA). The ND50 is typically0.05-0.2 µg/mL.
Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 μg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse CCL3/MIP‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-450-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CCL3/MIP‑1 alpha at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
CCL3/MIP‑1 alpha was detected in perfusion fixed frozen sections of mouse thymus using Goat Anti-Mouse CCL3/MIP‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog #AF-450-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.