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Human CCL3/CCL4 Antibody

Catalog #: MAB270
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
MAB270-SP
MAB270-100
MAB270-500
CCL3/CCL4 in U266 Human Cell Line.
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Citations (9)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CCL3/CCL4 Antibody Summary

Species Reactivity
Human
Specificity
Detects human CCL3/MIP-1 alpha in direct ELISAs and Western blots. In direct ELISAs, greater than 75% cross-reactivity with recombinant human (rh) CCL4/MIP-1 beta is observed while approximately 30% cross-reactivity with recombinant mouse (rm) CCL3, rmCCL4/MIP-1 beta, and rhCCL14 is observed. No cross-reactivity with rhCCL1, 2, 5, 7, 8, 11, 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, rmCCL1, 2, 5, 6, 7, 9/10/MIP-1 gamma, 11, 12, 17, 19, 20, 21, 22, 24, 25, or recombinant rat CCL20 is observed.
Source
Monoclonal Mouse IgG1 Clone # 93321
Purification
Protein A or G purified from hybridoma culture supernatant
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 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
Recombinant Human CCL3/MIP‑1 alpha isoform LD78a (Catalog # 270-LD) under non-reducing conditions only
Immunocytochemistry
8-25 µg/mL
Immersion fixed U266 human myeloma cell line
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 0.075-0.375 µg/mL in the presence of 10 ng/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.

Scientific Data

Immunocytochemistry View Larger

CCL3/CCL4 in U266 Human Cell Line. CCL3/CCL4 was detected in immersion fixed U266 human myeloma cell line (positive staining) and HL‑60 human acute promyelocytic leukemia cell line (negative staining) using Mouse Anti-Human CCL3/CCL4 Monoclonal Antibody (Catalog # MAB270) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutralization by Human CCL3/CCL4 Antibody. View Larger

Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutralization by Human CCL3/CCL4 Antibody. Recombinant Human CCL3/MIP-1a isoform LD78a (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-1a isoform LD78a (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CCL3/CCL4 Monoclonal Antibody (Catalog # MAB270). The ND50 is typically 0.075-0.375 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 0.5 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: CCL3/CCL4

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.
Long Name
CCL3/MIP-1 alpha and CCL4/MIP-1 beta Cross-reactive
Alternate Names
C-C motif chemokine;C-C motif chemokine 3;G0S19-1;LD78ALPHA;MIP1A;MIP-1-alpha;SCYA3; CCL3/CCL4

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Citations for Human CCL3/CCL4 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.

9 Citations: Showing 1 - 9
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  1. B cells inhibit bone formation in rheumatoid arthritis by suppressing osteoblast differentiation
    Authors: W Sun, N Meednu, A Rosenberg, J Rangel-Mor, V Wang, J Glanzman, T Owen, X Zhou, H Zhang, BF Boyce, JH Anolik, L Xing
    Nat Commun, 2018-12-03;9(1):5127.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  2. Distinct susceptibility of HIV vaccine vector-induced CD4 T cells to HIV infection
    Authors: S Auclair, F Liu, Q Niu, W Hou, G Churchyard, C Morgan, N Frahm, S Nitayaphan, P Pitisuthit, S Rerks-Ngar, JT Kimata, L Soong, G Franchini, M Robb, J Kim, N Michael, H Hu
    PLoS Pathog., 2018-02-23;14(2):e1006888.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. Elevated ACKR2 expression is a common feature of inflammatory arthropathies
    Authors: HM Baldwin, MD Singh, V Codullo, V King, H Wilson, I McInnes, GJ Graham
    Rheumatology (Oxford), 2017-09-01;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  4. An initial investigation into endothelial CC chemokine expression in the human rheumatoid synovium.
    Authors: Rump L, Mattey D, Kehoe O, Middleton J
    Cytokine, 2017-09-01;97(0):133-140.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  5. Macrophage inflammatory protein derivative ECI301 enhances the alarmin-associated abscopal benefits of tumor radiotherapy.
    Authors: Kanegasaki S, Matsushima K, Shiraishi K, Nakagawa K, Tsuchiya T
    Cancer Res, 2014-07-18;74(18):5070-8.
    Species: Mouse
    Sample Types: Beads
    Applications: Co-Immunoprecipitation
  6. Lymphatic specific disruption in the fine structure of heparan sulfate inhibits dendritic cell traffic and functional T cell responses in the lymph node.
    Authors: Yin X, Johns S, Kim D, Mikulski Z, Salanga C, Handel T, Macal M, Zuniga E, Fuster M
    J Immunol, 2014-02-03;192(5):2133-42.
    Species: Human
    Sample Types: Whole Cells
    Applications: Blocking
  7. A comprehensive ex vivo functional analysis of human NKT cells reveals production of MIP1-alpha and MIP1-beta, a lack of IL-17, and a Th1-bias in males.
    Authors: Snyder-Cappione JE, Tincati C, Eccles-James IG, Cappione AJ, Ndhlovu LC, Koth LL, Nixon DF
    PLoS ONE, 2010-11-03;5(11):e15412.
    Species: Human
    Sample Types: Whole Cells
    Applications: ELISpot Development
  8. Intratumoral induction of CD103 triggers tumor-specific CTL function and CCR5-dependent T-cell retention.
    Authors: Franciszkiewicz K, Le Floc'h A, Jalil A, Vigant F, Robert T, Vergnon I, Mackiewicz A, Benihoud K, Validire P, Chouaib S, Combadiere C, Mami-Chouaib F
    Cancer Res., 2009-07-28;69(15):6249-55.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  9. Induction of host chemotactic response by Encephalitozoon spp.
    Authors: Fischer J, West J, Agochukwu N, Suire C, Hale-Donze H
    Infect. Immun., 2006-12-18;75(4):1619-25.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization

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