|Detection of Mouse CCL3/MIP‑1 alpha by Western Blot. 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 in Mouse Thymus. 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.
Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutralization by Mouse CCL3/MIP‑1 alpha Antibody. Recombinant Mouse|
CCL3/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 typically
The macrophage inflammatory proteins 1 alpha and 1 beta, two closely related but distinct proteins, were originally co-purified from medium conditioned by a LPS-stimulated murine macrophage cell line. Mature mouse MIP-1 alpha shares approximately 77% and 70% amino acid identity with human MIP-1 alpha and mouse MIP-1 beta, respectively. MIP‑1 proteins are expressed primarily in T cells, B cells, and monocytes after antigen or mitogen stimulation. The MIP-1 proteins are members of the beta (C-C) subfamily of chemokines.
Both MIP-1 alpha and MIP-1 beta are monocyte chemoattractants in vitro. Additionally, the MIP-1 proteins have been reported to have chemoattractant and adhesive effects on lymphocytes, with MIP-1 alpha and MIP-1 beta preferentially attracting CD8+ and CD4+ T cells, respectively. MIP-1 alpha has also been shown to attract B cells as well as eosinophils. MIP-1 proteins have been reported to have multiple effects on hematopoietic precursor cells and MIP-1 alpha has been identified as a stem cell inhibitory factor that can inhibit the proliferation of hematopoietic stem cells in vitro as well as in vivo. In the same assays, MIP-1 beta was reported to be much less active. The functional receptor for MIP-1 alpha has been identified as CCR1 and CCR5.
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