Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Rat

Cited:

Rat

Applications

Validated:

Western Blot, Neutralization

Cited:

Immunohistochemistry, Neutralization, In vivo assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 769324
View all CCL3/MIP-1 alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant rat CCL3/MIP‑1 alpha
Ala24-Ala92
Accession # P50229

Specificity

Detects rat CCL3/MIP‑1 alpha in direct ELISAs.

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 Rat CCL3/MIP-1 alpha Antibody (769324)

Detection of Rat CCL3/MIP-1a antibody by Western Blot.

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

Western blot shows lysates of NR8383 rat alveolar macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 4 hours. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Rat CCL3/MIP-1a Monoclonal Antibody (Catalog # MAB66252) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for CCL3/MIP-1a at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutral-ization by Rat CCL3/ MIP‑1 alpha Antibody.

Chemotaxis Induced by CCL3/MIP‑1 alpha and Neutral-ization by Rat CCL3/ MIP‑1 alpha Antibody.

Recombinant Rat CCL3/MIP-1a (Catalog # 6625-MA) induces chemotaxis in the BaF3 mouse pro-B cell line transfected with human CCR5 in a dose-dependent manner (orange line). Chemotaxis elicited by Recombinant Rat CCL3/MIP-1a (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Rat CCL3/MIP-1a Monoclonal Antibody (Catalog # MAB66252). The ND50 is typically 0.075-0.45 µg/mL.
Detection of CCL3/MIP-1 alpha by Immunocytochemistry/ Immunofluorescence

Detection of CCL3/MIP-1 alpha by Immunocytochemistry/ Immunofluorescence

Effect of neutralizing antibodies for chemokines/chemoattractans in monocytes/MΦs in vitro cell migration assays. (A) The effect of the neutralizing antibodies against CCL2 (500 ng/mL), CCL3 (500 ng/mL), and CCL21 (500 ng/mL) in the monocytes/MΦs migration assay under the chemoattractant effect of the CM of the glomeruli from DN rats. HAM-F10 and 5% FBS were used as negative and positive control, respectively, of in vitro stimuli for migration. DAPI was used to stain the nuclei of monocytes/MΦs. Representative quadrants show monocytes/MΦs in blue (400× magnification). (B) Quantification of positive intraglomerular area (%) of deconvoluted color for DAB in (A). Graphs represent distribution of each sample and the mean ± S.D. * p < 0.05, and *** p < 0.001 versus Ctrl rats. #p < 0.05 and ###p < 0.001, DN + MRS1754 versus DN rats. n = 4. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37446007), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Rat CCL3/MIP-1 alpha Antibody (769324)

Application
Recommended Usage

Western Blot

2 µg/mL
Sample: NR8383 rat alveolar macrophage cell line treated with LPS

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 0.075-0.45 μg/mL in the presence of 10 ng/mL Recombinant Rat CCL3/MIP‑1 alpha.

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.

Calculators

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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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 rat CCL3 shares 74%, 91%, and 88% amino acid sequence identity with human, mouse, 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

  1. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
  2. Shi, M.M. et al. (1995) Biochem. Biophys. Res. Commun. 211:289.
  3. Graham, G.J. et al. (1990) Nature 344:442.
  4. Wagner, L. et al. (1998) Nature 391:908.
  5. Graham, G.J. et al. (1994) J. Biol. Chem. 269:4974.
  6. Ren, M. et al. (2010) EMBO J. 29:3952.
  7. Weber, M. et al. (2004) Mol. Biol. Cell 15:2492
  8. Taub, D.D. et al. (1993) Science 260:355.
  9. Bernardini, G. et al. (2008) Blood 111:3626.
  10. Lee, S.C. et al. (2000) J. Immunol. 164:3392.
  11. Wu, Y. et al. (2008) J. Immunol. 181:6384.
  12. Cook, D.N. et al. (1995) Science 269:1583.
  13. Chintalacharuvu, S.R. et al. (2005) Immunol. Lett. 100:202.
  14. Ajuebor, M.N. et al. (2004) Eur. J. Immunol. 34:2907.
  15. Cocchi, F. et al. (1995) Science 270:1811.
  16. 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

P50229

Additional CCL3/MIP-1 alpha Products

Product Documents for Rat CCL3/MIP-1 alpha Antibody (769324)

Certificate of Analysis

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Product Specific Notices for Rat CCL3/MIP-1 alpha Antibody (769324)

For research use only

Citations for Rat CCL3/MIP-1 alpha Antibody (769324)

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