Human CCL3/MIP-1 alpha Antibody AF-270-NA: R&D Systems

Human CCL3/MIP-1 alpha Antibody

(11 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human CCL3/MIP‑1 alpha  in ELISAs and Western blots. In sandwich immunoassays, less than 0.05% cross-reactivity with recombinant human (rh) MIP-1 beta, rhMIP-1δ, rhMIP-3 alpha, rhMIP-3 beta, and recombinant mouse (rm) MIP-1 alpha is observed. Neutralizes the biological activity of recombinant human CCL3/MIP‑1 alpha. In the chemotaxis assay this antibody will also partially neutralize the biological activity of rhMIP-1 beta at a 20 fold higher IgG concentration. It will not neutralize the biological activity of rmMIP-1 beta or rmMIP-1 alpha.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human CCL3/MIP-1 alpha
    Ala27-Ala92
    Accession # P10147
  • 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 CCL3/MIP‑1 alpha isoform LD78a (Catalog # 270-LD)
  • Immunohistochemistry
    5-15 µg/mL
    See below
    • Human CCL3/MIP-1 alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human CCL3/MIP‑1 alpha Antibody (Catalog # AF-270-NA)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human CCL3/MIP‑1 alpha Biotinylated Antibody (Catalog # BAF270)
  • ELISA Standard
     
    Recombinant Human CCL3/MIP-1 alpha Protein (Catalog # 270-LD)
  • Neutralization
    Measured by its ability to neutralize CCL3/MIP‑1 alpha -induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 2-8 µg/mL in the presence of 0.1 µg/mL Recombinant Human 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.
Data Examples

Chemotaxis Induced by CCL3/MIP‑1 alpha  and Neutral­ization by Human CCL3/
MIP‑1 alpha Antibody.
Recombinant Human CCL3/MIP‑1 alpha (Catalog # 270-LD) 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
CCL3/MIP‑1 alpha (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL3/MIP‑1 alpha Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF-270-NA). The ND50 is typically 2-8 µg/mL.

CCL3/MIP‑1 alpha in Human Tonsil. CCL3/MIP‑1 alpha was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CCL3/MIP‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-270-NA) at 10 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 in lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

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: CCL3/MIP-1 alpha

The macrophage inflammatory proteins -1 alpha and -1 beta were originally co-purified from medium conditioned by an LPS-stimulated murine macrophage cell line. Human MIP-1 alpha refers to the products of several independently cloned cDNAs, including LD78, pL78, pAT464, and GOS19. These cDNAs all code for the same human protein that is a homologue of the murine MIP-1 alpha. Mature MIP-1 alpha and MIP-1 beta in both human and mouse share approximately 70% homology at the amino acid level. 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. The functional receptor for MIP-1 alpha has been identified as CCR1 and CCR5.

  • References:
    1. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
  • Entrez Gene IDs:
    6348 (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine)
  • Alternate Names:
    C-C motif chemokine 3; MIP1-(a); AI323804; CCL3; chemokine (C-C motif) ligand 3; G0S19-1; LD78a; LD78alpha; MIP1 alpha; MIP-1 alpha; Mip1a; MIP-1alpha; MIP1-alpha; Scya3
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.

11 Citations: Showing 1 - 10
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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. Noninvasive detection of acute and chronic injuries in human renal transplant by elevation of multiple cytokines/chemokines in urine.
    Authors: Hu H, Kwun J, Aizenstein BD, Knechtle SJ
    Transplantation, 2009;87(12):1814-20.
    Species: Human
    Sample Type: Urine
    Application: Antibody Array Development
  3. Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin.
    Authors: Wu W, Mehta H, Chakrabarty K, Booth JL, Duggan ES, Patel KB, Ballard JD, Coggeshall KM, Metcalf JP
    J. Immunol., 2009;183(9):5799-806.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  4. Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo.
    Authors: Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB
    AIDS Res. Hum. Retroviruses, 2008;24(3):453-62.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  5. Effect of serum content and diluent selection on assay sensitivity and signal intensity in multiplex bead-based immunoassays.
    Authors: Pfleger C, Schloot N, ter Veld F
    J. Immunol. Methods, 2007;329(1):214-8.
    Species: Human
    Sample Type: Serum
    Application: Luminex Development
  6. Fluorescence single-molecule counting assays for high-sensitivity detection of cytokines and chemokines.
    Authors: Qui H, Ferrell EP, Nolan N, Phelps BH, Tabibiazar R, Whitney DH, Naelfski EA
    Clin. Chem., 2007;53(11):2010-2.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  7. HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1.
    Authors: Grivel JC, Elliott J, Lisco A, Biancotto A, Condack C, Shattock RJ, McGowan I, Margolis L, Anton P
    AIDS, 2007;21(10):1263-72.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  8. Abnormal activation and cytokine spectra in lymph nodes of people chronically infected with HIV-1.
    Authors: Biancotto A, Grivel JC, Iglehart SJ, Vanpouille C, Lisco A, Sieg SF, Debernardo R, Garate K, Rodriguez B, Margolis LB, Lederman MM
    Blood, 2007;109(10):4272-9.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Luminex Development
  9. MIP-1alpha (CCL3) is a downstream target of FGFR3 and RAS-MAPK signaling in multiple myeloma.
    Authors: Masih-Khan E, Trudel S, Heise C, Li Z, Paterson J, Nadeem V, Wei E, Roodman D, Claudio JO, Bergsagel PL, Stewart AK
    Blood, 2006;108(10):3465-71.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
  10. Gene copy number regulates the production of the human chemokine CCL3-L1.
    Authors: Townson JR, Barcellos LF, Nibbs RJ
    Eur. J. Immunol., 2002;32(10):3016-26.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  11. 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
Expand to show all 11 Citations
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