Detects mouse CCL3/MIP-1 alpha in direct ELISAs and Western blots. In Western blots, less than 5% cross-reactivity with recombinant human (rh) MIP-1 beta, rhMIP-1 alpha, and recombinant mouse MIP-1 beta is observed.
Measured by its ability to neutralize CCL3/MIP‑1 alpha -induced chemotaxis in BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.3-1 µg/mL in the presence of 10 ng/mL Recombinant Mouse CCL3/MIP‑1 alpha Isoform LD78a.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
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 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 Polyclonal Antibody (Catalog # AB-450-NA). The ND50 is typically 0.3-1 µg/mL.
Preparation and Storage
Reconstitute at 1 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: CCL3/MIP-1 alpha
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.
Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
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