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Human CCL3/MIP-1 alpha Antibody

R&D Systems, part of Bio-Techne | Catalog # AF-270-NA

R&D Systems, part of Bio-Techne
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AF-270-NA
AF-270-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

ELISA Capture (Matched Antibody Pair), Immunohistochemistry, Neutralization, Western Blot

Cited:

Antibody Array Development, ELISA Development, Flow Cytometry, Luminex Development, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant human CCL3/MIP-1 alpha
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.

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-1a antibody in Human Tonsil by Immunohistochemistry (IHC-P).

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.

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

Western Blot

0.1 µg/mL
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

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human CCL3/MIP-1 alpha Biotinylated Antibody (Catalog # BAF270)
  • Standard: Recombinant Human CCL3/MIP-1 alpha Protein (Catalog # 270-LD)
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.

Reconstitution Buffer Available:
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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

  1. 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

6348 (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine); 102136134 (Cynomolgus Monkey)

Gene Symbol

CCL3

UniProt

Additional CCL3/MIP-1 alpha Products

Product Documents for Human CCL3/MIP-1 alpha Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CCL3/MIP-1 alpha Antibody

For research use only

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