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Mouse CCL4/MIP-1 beta Antibody

Catalog #: AF-451-NA
13 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF-451-NA
AF-451-SP
Detection of Mouse CCL4/MIP‑1 beta by Western Blot.
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Citations (13)
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Mouse CCL4/MIP-1 beta Antibody Summary

Species Reactivity
Mouse
Specificity
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 alpha is observed. 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.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant mouse CCL4/MIP-1 beta
Ala24-Asn92
Accession # Q5QNV9
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunohistochemistry
1-15 µg/mL
See below
Immunocytochemistry
1-15 µg/mL
See below
Neutralization
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.

Scientific Data

Western Blot Detection of Mouse CCL4/MIP-1 beta antibody by Western Blot. View Larger

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.

Immunocytochemistry CCL4/MIP-1 beta antibody in RAW 264 by Immunocytochemistry (ICC).7 Mouse Cell Line by Immunocytochemistry (ICC). View Larger

CCL4/MIP‑1 beta in RAW 264.7 Mouse Cell Line. CCL4/MIP-1 beta was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line untreated (right panel) and treated with LPS (left panel) using Goat Anti-Mouse CCL4/MIP-1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-451-NA) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry CCL4/MIP-1 beta antibody in Mouse Small Intestine by Immunohistochemistry (IHC-Fr). View Larger

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.

Neutralization Chemotaxis Induced by CCL4/MIP‑1 beta and Neutral-ization by Mouse CCL4/ MIP‑1 beta Antibody. View Larger

Chemotaxis Induced by CCL4/MIP‑1 beta and Neutral-ization 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
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.

References
  1. Rot, A. and U.H. von Andrian (2004) Annu. Rev. Immunol. 22:891.
  2. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
  3. Sun, X. et al. (2006) Infec. Immun. 74:5943.
  4. Bisset, L.R. and Schmid-Grendelmeier, P. (2005) Curr. Opin. Pulm. Med. 11:35.
  5. Frangogiannis, N.G. (2004) Inflamm. Res. 53:585.
  6. Bystry, R.S. et al. (2001) Nat. Immunol. 2:1126.
  7. Oliveira, S.H.P. et al. (2002) J. Leukoc. Biol. 71:1019.
  8. Schall, T.J. et al. (1993) J. Exp. Med. 177:1821.
  9. Guan, E. et al. (2001) J. Biol. Chem. 276:12404.
  10. Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
  11. Guan, E. et al. (2004) J. Cell. Biochem. 92:53.
Entrez Gene IDs
6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine); 102117861 (Cynomolgus Monkey)
Alternate Names
ACT2; ACT-2; AT744.1; C-C motif chemokine 4; CCL4; chemokine (C-C motif) ligand 4; Exodus-3; G-26 T-lymphocyte-secreted protein; G-26; HC21; LAG1; LAG-1; Lymphocyte activation gene 1 protein; Macrophage inflammatory protein 1-beta; MIP1 beta; MIP-1 beta; MIP1B; MIP1B1; MIP-1-beta; MIP-1-beta(1-69); PAT 744; Protein H400; SCYA2; SCYA4; secreted protein G-26; SIS-gamma; small inducible cytokine A4 (homologous to mouse Mip-1b); Small-inducible cytokine A4; T-cell activation protein 2

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Citations for Mouse CCL4/MIP-1 beta Antibody

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.

13 Citations: Showing 1 - 10
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  1. IL9 Polarizes Macrophages to M1 and Induces the Infiltration of Antitumor Immune Cells via MIP-1 and CXCR3 Chemokines
    Authors: Van Anh Do-Thi, Sang Min Park, Song Mi Park, Hye Jin Jeong, Geunyoung Cho, Hyun-Jung An et al.
    Cancer Research Communications
  2. Gestational Bisphenol A Exposure Impacts Embryonic Hypothalamic Microglia Numbers, Ramification, and Phagocytic Cups
    Authors: Jessica M. Rosin, Nikol Tretiakov, Emily Hanniman, Kiana Hampton, Deborah M. Kurrasch
    Frontiers in Neuroscience
  3. Radiation Triggers a Dynamic Sequence of Transient Microglial Alterations in Juvenile Brain
    Authors: AM Osman, Y Sun, TC Burns, L He, N Kee, N Oliva-Vila, A Alevyzaki, K Zhou, L Louhivuori, P Uhlén, E Hedlund, C Betsholtz, VM Lauschke, J Kele, K Blomgren
    Cell Rep, 2020-06-02;31(9):107699.
    Species: Mouse
    Sample Types:
    Applications: IHC
  4. Recruitment of CD103+ dendritic cells via tumor-targeted chemokine delivery enhances efficacy of checkpoint inhibitor immunotherapy
    Authors: JM Williford, J Ishihara, A Ishihara, A Mansurov, P Hosseinchi, TM Marchell, L Potin, MA Swartz, JA Hubbell
    Sci Adv, 2019-12-11;5(12):eaay1357.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  5. Single-Cell RNA Sequencing of Microglia throughout the Mouse Lifespan and in the Injured Brain Reveals Complex Cell-State Changes
    Authors: Timothy R. Hammond, Connor Dufort, Lasse Dissing-Olesen, Stefanie Giera, Adam Young, Alec Wysoker et al.
    Immunity
  6. IFN? Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury
    Authors: VE Thaney, AM O'Neill, MM Hoefer, R Maung, AB Sanchez, M Kaul
    Sci Rep, 2017-04-20;7(0):46514.
    Species: Mouse, Rat
    Sample Types: Whole Cells
    Applications: Neutralization
  7. Identification of Chemoattractive Factors Involved in the Migration of Bone Marrow-Derived Mesenchymal Stem Cells to Brain Lesions Caused by Prions
    Authors: Chang-Hyun Song, Osamu Honmou, Hidefumi Furuoka, Motohiro Horiuchi
    Journal of Virology
  8. Comprehensive assessment of chemokine expression profiles by flow cytometry.
    Authors: Eberlein J, Nguyen TT, Victorino F, Golden-Mason L, Rosen HR, Homann D
    J. Clin. Invest., 2010-02-08;120(3):907-23.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  9. CCL3-CCR5 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process.
    Authors: Wu Y, Li YY, Matsushima K, Baba T, Mukaida N
    J. Immunol., 2008-11-01;181(9):6384-93.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  10. Role of the chemokine decoy receptor D6 in balancing inflammation, immune activation, and antimicrobial resistance in Mycobacterium tuberculosis infection.
    Authors: Di Liberto D, Locati M, Caccamo N, Vecchi A, Meraviglia S, Salerno A, Sireci G, Nebuloni M, Caceres N, Cardona PJ, Dieli F, Mantovani A
    J. Exp. Med., 2008-08-11;205(9):2075-84.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  11. CCL4 protects from type 1 diabetes by altering islet beta-cell-targeted inflammatory responses.
    Authors: Meagher C, Arreaza G, Peters A, Strathdee CA, Gilbert PA, Mi QS, Santamaria P, Dekaban GA, Delovitch TL
    Diabetes, 2007-03-01;56(3):809-17.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  12. CC chemokines mediate leukocyte trafficking into the central nervous system during murine neurocysticercosis: role of gamma delta T cells in amplification of the host immune response.
    Authors: Cardona AE, Gonzalez PA, Teale JM
    Infect. Immun., 2003-05-01;71(5):2634-42.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  13. Role of CC chemokines (macrophage inflammatory protein-1 beta, monocyte chemoattractant protein-1, RANTES) in acute lung injury in rats.
    Authors: Bless NM, Huber-Lang M, Guo RF, Warner RL, Schmal H, Czermak BJ, Shanley TP, Crouch LD, Lentsch AB, Sarma V, Mulligan MS, Friedl HP, Ward PA
    J. Immunol., 2000-03-01;164(5):2650-9.
    Species: Rat
    Sample Types: BALF
    Applications: ELISA Development

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