Detects CCL4/MIP-1 beta in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) MIP‑1 beta and recombinant mouse (rm) MIP-1 gamma is observed and approximately 10% cross-reactivity with rmMIP-1 alpha is observed. Neutralizes mouse MIP-1 beta. This antibody will not neutralize the biological activity of rmMIP-1 alpha. In the chemotaxis assay this antibody will partially neutralize rhMIP-1 alpha and rhMIP-1 beta at a 10‑30 fold higher IgG concentration.
Polyclonal Goat IgG
E. coli-derived recombinant mouse CCL4/MIP-1 beta Ala24-Asn92 Accession # Q5QNV9
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize CCL4/MIP‑1 beta -induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.2-1.0 µg/mL in the presence of 25 ng/mL Recombinant Mouse CCL4/MIP‑1 beta.
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.
Detection of Mouse CCL4/MIP‑1 beta 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 CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-451-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CCL4/MIP‑1 beta at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
CCL4/MIP‑1 beta in Mouse Small Intestine. CCL4/MIP‑1 beta was detected in perfusion fixed frozen sections of mouse small intestine using Goat Anti-Mouse CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-451-NA) at 1.7 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 Peyer's patches. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Chemotaxis Induced by CCL4/MIP‑1 beta and Neutralization by Mouse CCL4/ MIP‑1 beta Antibody. Recombinant Mouse CCL4/MIP‑1 beta (Catalog # 451‑MB) 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 CCL4/ MIP‑1 beta (25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-451-NA). The ND50 is typically 0.2‑1.0 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: CCL4/MIP-1 beta
CCL4, also known as macrophage inflammatory protein 1 beta (MIP-1 beta ), is a 12 kDa beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells (1, 2). CCL4 attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammation such as allergic asthma and ischemic myocardium (3-8). A CCL4 deficiency in mice promotes the development of autoantibodies, possibly as a result of compromised regulatory T cell recruitment (6). CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha (9). The first two N-terminal amino acids can be cleaved from human CCL4 by CD26/DPPIV (10, 11). Both the full length and truncated forms exert biological activity through CCR5, and the truncated form additionally interacts with CCR1 and CCR2 (10). In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor (2). Both forms of CCL4 block HIV-1 infection of T cells by inducing the downregulation of CCR5 (10). Mature mouse CCL4 shares 77% and 86% aa sequence identity with human and rat CCL4, respectively.
Rot, A. and U.H. von Andrian (2004) Annu. Rev. Immunol. 22:891.
Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
Sun, X. et al. (2006) Infec. Immun. 74:5943.
Bisset, L.R. and Schmid-Grendelmeier, P. (2005) Curr. Opin. Pulm. Med. 11:35.
Frangogiannis, N.G. (2004) Inflamm. Res. 53:585.
Bystry, R.S. et al. (2001) Nat. Immunol. 2:1126.
Oliveira, S.H.P. et al. (2002) J. Leukoc. Biol. 71:1019.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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