Human CCL3/MIP-1 alpha Antibody
R&D Systems, part of Bio-Techne | Catalog # AF-270-NA
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Conjugate
Catalog #
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization
Cited:
Western Blot, Neutralization, Flow Cytometry, Antibody Array Development, ELISA Development, Functional Assay, Luminex Development
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
E. coli-derived recombinant human CCL3/MIP-1 alpha
Ala27-Ala92
Accession # P10147
Ala27-Ala92
Accession # P10147
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-1 delta, 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.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human CCL3/MIP-1 alpha Antibody
Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutral-ization by Human CCL3/ MIP‑1 alpha Antibody.
Recombinant Human CCL3/MIP-1a (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 (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CCL3/MIP-1a 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-1a was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CCL3/MIP-1a 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.
Human CCL3 / MIP-1 alpha ELISA Standard Curve
Recombinant Human CCL3/MIP-1 alpha Isoform LD78a (Catalog # 270-LD) was serially diluted and captured by Goat Anti-Human CCL3/MIP-1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-270-NA) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human CCL3/MIP-1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-270-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)Applications for Human CCL3/MIP-1 alpha Antibody
Application
Recommended Usage
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil
Sample: Immersion fixed paraffin-embedded sections of human tonsil
Western Blot
0.1 µg/mL
Sample: Recombinant Human CCL3/MIP-1 alpha isoform LD78a (Catalog # 270-LD)
Sample: Recombinant Human CCL3/MIP-1 alpha isoform LD78a (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.
Human CCL3/MIP-1 alpha Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 2 reviews rated 5 using AF-270-NA in the following applications:
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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
- Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
Alternate Names
G0S19-1, LD78a, MIP-1 alpha, MIP1 alpha, MIP1A, PAT 464.1, SCYA3, SIS-beta
Entrez Gene IDs
Gene Symbol
CCL3
UniProt
Additional CCL3/MIP-1 alpha Products
Product Documents for Human CCL3/MIP-1 alpha Antibody
Product Specific Notices for Human CCL3/MIP-1 alpha Antibody
For research use only
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