Detects mouse CCL3/MIP‑1 alpha in direct ELISAs and Western blots. In direct ELISAs, 100% cross-reactivity with recombinant rat CCL3 and 50% cross-reactivity with recombinant human CCL3 is observed. No cross-reactivity with recombinant canine CCL3, recombinant cotton rat CCL3, recombinant mouse (rm) CCL4/MIP-1 beta, or rmCCL5 is observed.

Clonality

Monoclonal

Host

Rat

Isotype

IgG_2B

Scientific Data Images for Mouse CCL3/MIP-1 alpha Antibody

Detection of Mouse CCL3/MIP‑1 alpha by Western Blot.

Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Rat Anti-Mouse CCL3/MIP-1a Monoclonal Antibody (Catalog # MAB4501) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for CCL3/MIP-1a at approximately 8 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

CCL3/MIP‑1 alpha in RAW 264.7 Mouse Cell Line.

CCL3/MIP-1a was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line stimulated (upper panel) or not (lower panel) with LPS using Rat Anti-Mouse CCL3/MIP-1a Monoclonal Antibody (Catalog # MAB4501) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm of stimulated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Mouse CCL3/MIP-1 alpha Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed RAW 264.7 mouse monocyte/macrophage cell line stimulated with LPS

Western Blot

1 µg/mL
Sample: RAW 264.7 mouse monocyte/macrophage cell line treated with LPS

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. 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

CCL3, also known as macrophage inflammatory protein 1 alpha (MIP-1 alpha) and LD78, is a member of the beta or CC subfamily of chemokines and is closely related to CCL4/MIP-1 beta. Chemokines comprise a large family of small secreted proteins that are involved in immune and inflammatory responses. CCL3 expression can be induced in a variety of hematopoietic cells, fibroblasts, smooth muscle cells, and epithelial cells (1). Mature mouse CCL3 shares 73%, 91%, and 82% amino acid sequence identity with human, rat, and cotton rat CCL3, respectively (2). CCL3 is an approximately 8 kDa chemokine that forms complexes with sulfated proteoglycans (3, 4). In a reversible process, CCL3 associates into noncovalently-linked dimers which then form tetramers and high molecular weight polymers (5, 6). These complexes of CCL3 are protected from proteolytic digestion by insulin degrading enzyme (IDE) which can cleave the monomeric chemokine (6). CCL3 exerts its biological functions through interactions with CCR1, CCR3, and CCR5 (1). It is cleared from the extracellular space by internalization via the decoy chemokine receptor D6 (7). CCL3 promotes the chemoattraction, adhesion to activated vascular endothelium, and cellular activation of many hematopoietic cell types including activated T cells, NK cells, neutrophils, monocytes, immature dendritic cells, and eosinophils (1, 8-10). CCL3 is also known as stem cell inhibitor (SCI) and can inhibit the proliferation of hematopoietic progenitor cells (3). CCL3 bioactivity contributes to tumor metastasis and the inflammatory components of viral infection, rheumatoid arthritis, and hepatitis (11-14), although it also can suppress the replication of HIV (15). CCL3 additionally promotes hyperalgesia by sensitizing sensory neurons to TRPV1-mediated noxious stimulation (16).

References

Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
Davatelis, G. et al. (1988) J. Exp. Med. 167:1939.
Graham, G.J. et al. (1990) Nature 344:442.
Wagner, L. et al. (1998) Nature 391:908.
Graham, G.J. et al. (1994) J. Biol. Chem. 269:4974.
Ren, M. et al. (2010) EMBO J. 29:3952.
Weber, M. et al. (2004) Mol. Biol. Cell 15:2492
Taub, D.D. et al. (1993) Science 260:355.
Bernardini, G. et al. (2008) Blood 111:3626.
Lee, S.C. et al. (2000) J. Immunol. 164:3392.
Wu, Y. et al. (2008) J. Immunol. 181:6384.
Cook, D.N. et al. (1995) Science 269:1583.
Chintalacharuvu, S.R. et al. (2005) Immunol. Lett. 100:202.
Ajuebor, M.N. et al. (2004) Eur. J. Immunol. 34:2907.
Cocchi, F. et al. (1995) Science 270:1811.
Zhang, N. et al. (2005) Proc. Natl. Acad. Sci. 102:4536.

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

P10855

Additional CCL3/MIP-1 alpha Products

Product Documents for Mouse CCL3/MIP-1 alpha Antibody

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COA

SDS

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

For research use only

Citations
Reviews

Mouse CCL3/MIP-1 alpha Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB4501

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications