Immunohistochemistry, Immunohistochemistry-Frozen, Neutralization, Immunocytochemistry, Bioassay, Blocking, Co-Immunoprecipitation, ELISpot Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # 93321

View all CCL3/CCL4 Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CCL3/MIP-1 alpha
Ala27-Ala92
Accession # P10147

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human CCL3/CCL4 Antibody

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.

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.

Applications for Human CCL3/CCL4 Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed U266 human myeloma cell line

Western Blot

1 µg/mL
Sample: Recombinant Human CCL3/MIP‑1 alpha isoform LD78a (Catalog # 270-LD) under non-reducing conditions only

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.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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/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

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

UniProt

P10147

Additional CCL3/CCL4 Products

Product Documents for Human CCL3/CCL4 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human CCL3/CCL4 Antibody

For research use only

Citations
Reviews

Human CCL3/CCL4 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB270

Key Product Details

Species Reactivity

Validated:

Cited:

Applications