Human CCL4/MIP-1 beta Antibody

(7 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CCL4/MIP-1 beta in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with  recombinant mouse (rm) MIP-1 alpha is observed, approximately 20% cross-reactivity with recombinant human (rh) MIP-1 alpha and recombinant cotton rat MIP-1 alpha is observed, and less than 1% cross-reactivity with rhMCP-2, rmMIP-1 beta, rmMCP-2, and recombinant viral MIP-1 beta  is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CCL4/MIP-1 beta
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human CCL4/MIP‑1 beta (Catalog # 271-BME)
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 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 0.3-1.5 µg/mL in the presence of 40 ng/mL Recombinant Human 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.
Data Examples
Chemotaxis Induced by CCL4/MIP‑1 beta  and Neutral­ization by Human CCL4/MIP‑1 beta  Antibody.

Recombinant Human CCL4/MIP‑1 beta (Catalog # 271-BME) 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 Human CCL4/MIP‑1 beta (40 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-271-NA). The ND50 is typically 0.3-1.5 µg/mL.

Immunohistochemistry
CCL4/MIP‑1 beta  in Human Alzheimer's Disease Brain.

CCL4/MIP‑1 beta was detected in immersion fixed paraffin-embedded sections of human Alzheimer's disease brain (hippocampus) using 15 µg/mL Goat Anti-Human CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑271‑NA) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-AEC Cell & Tissue Staining Kit (red; Catalog # CTS009) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunocytochemistry
CCL4/MIP‑1 beta  in Human PBMCs.

CCL4/MIP‑1 beta was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PHA and monensin using Goat Anti-Human CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-271-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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/MIP-1 beta is a beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells. It 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. CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha. It signals through CCR5, and an N-terminally trimmed form additionally interacts with CCR1 and CCR2. In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor.
  • Entrez Gene IDs:
    6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine)
  • 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
Related Research Areas
Citations:

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.

7 Citations: Showing 1 - 7
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Species
Applications
Sample Type
  1. Increased Levels of Macrophage Inflammatory Proteins Result in Resistance to R5-Tropic HIV-1 in a Subset of Elite Controllers.
    Authors: Walker W, Kurscheid S, Joshi S, Lopez C, Goh G, Choi M, Barakat L, Francis J, Fisher A, Kozal M, Zapata H, Shaw A, Lifton R, Sutton R, Fikrig E
    J Virol, 2015;89(10):5502-14.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. NK cell responses to simian immunodeficiency virus vaginal exposure in naive and vaccinated rhesus macaques.
    Authors: Shang L, Smith A, Duan L, Perkey K, Qu L, Wietgrefe S, Zupancic M, Southern P, Masek-Hammerman K, Reeves R, Johnson R, Haase A
    J Immunol, 2014;193(1):277-84.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Tissue
    Application: IHC
  3. A novel p38-MAPK signaling axis modulates neutrophil biology in head and neck cancer.
    Authors: Dumitru CA, Fechner MK, Hoffmann TK
    J. Leukoc. Biol., 2012;91(4):591-8.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  4. Villitis of unknown etiology is associated with a distinct pattern of chemokine up-regulation in the feto-maternal and placental compartments: implications for conjoint maternal allograft rejection and maternal anti-fetal graft-versus-host disease.
    Authors: Kim MJ, Romero R, Kim CJ, Tarca AL, Chhauy S, LaJeunesse C, Lee DC, Draghici S, Gotsch F, Kusanovic JP, Hassan SS, Kim JS
    J. Immunol., 2009;182(6):3919-27.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  5. Intestinal neuro-epithelial interactions modulate neuronal chemokines production.
    Authors: Tixier E, Galmiche JP, Neunlist M
    Biochem. Biophys. Res. Commun., 2006;344(2):554-61.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  6. The beta chemokines CCL4 and CCL5 enhance adhesion of specific CD4+ T cell subsets to human brain endothelial cells.
    Authors: Quandt J, Dorovini-Zis K
    J. Neuropathol. Exp. Neurol., 2004;63(4):350-62.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  7. Simian immunodeficiency viruses with defective nef genes show increased susceptibility to the noncytotoxic antiviral activity of CD8+ lymphocytes.
    Authors: Binninger-Schinzel D, Norley S, Adler HS, Oberg HH, Kurth R
    Virology, 2002;294(1):209-21.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
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