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Mouse CCL5/RANTES Antibody

R&D Systems, part of Bio-Techne | Catalog # AF478

R&D Systems, part of Bio-Techne
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AF478
AF478-SP

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunocytochemistry, Neutralization, Western Blot

Cited:

Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant mouse CCL5/RANTES

Specificity

Detects mouse CCL5/RANTES in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.30 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse CCL5/RANTES Antibody

Chemotaxis Induced by CCL5/RANTES and Neutralization by Mouse CCL5/RANTES Antibody.

Chemotaxis Induced by CCL5/RANTES and Neutralization by Mouse CCL5/RANTES Antibody.

Recombinant Mouse CCL5/RANTES (478-MR) 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 (AR002). Chemotaxis elicited by Recombinant Mouse CCL5/RANTES (0.025 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CCL5/ RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF478). The ND50 is typically 0.07-0.3 µg/mL.
CCL5/RANTES antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

CCL5/RANTES in Mouse Splenocytes.

CCL5/RANTES was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse CCL5/RANTES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF478) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Mouse CCL5/RANTES Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse CCL5/RANTES (Catalog # 478-MR)

Neutralization

Measured by its ability to neutralize CCL5/RANTES-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CCR5. The Neutralization Dose (ND50) is typically 0.07-0.3 µg/mL in the presence of 0.025 µg/mL Recombinant Mouse CCL5/RANTES.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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: CCL5/RANTES

CCL5, also known as RANTES (Regulated upon Activation, Normal T cell Expressed and presumably Secreted), is an 8 kDa beta-chemokine that plays a primary role in the inflammatory immune response by means of its ability to attract and activate leukocytes (1-3). Human and mouse RANTES exhibit cross-species activity on human and mouse cells (4). Mature mouse CCL5 shares 100% aa sequence identity with rat CCL5 and 75-88% with canine, cotton rat, feline, and human CCL5 (5). CCL5 is secreted by many cell types at inflammatory sites, and it exerts a wide range of activities through the receptors CCR1, CCR3, CCR4, and CCR5 (6, 7). Inflammatory responses can be impaired by the sequestration of CCL5 by the cytomegalovirus protein US28 (8). In humans, CCR5 binding to CCL5 inhibits the infectivity of R5 (M-tropic) but not X4 (T-tropic) strains of HIV-1 (9). The two N-terminal residues of CCL5 can be removed by CD26/DPPIV, generating a protein that functions as a chemotaxis inhibitor and more effectively blocks M-tropic HIV-1 infection of monocytes (10). Oligomerization of CCL5 on glycosaminoglycans is required for CCR1-mediated leukocyte adhesion and activation as well as CCL5’s interaction with the chemokine CXCL4/PF4 (11-13). The deposition of CCL5 on activated vascular endothelial cells is crucial for monocyte adhesion to damaged vasculature, but CCL5 oligomerization is not required for the extravasation of adherent leukocytes (14-16). CCL5 is upregulated in breast cancer and promotes tumor progression through the attraction of pro-inflammatory macrophages in addition to its actions on tumor cells, stromal cells, and the vasculature (17).

References

  1. Schall, T.J. et al. (1990) Nature 347:669.
  2. Bacon, K.B. et al. (1995) Science 269:1727.
  3. Fischer, F.R. et al. (2001) J. Immunol. 167:1637.
  4. Schall, T.J. et al. (1992) Eur. J. Immunol. 22:1477.
  5. Heeger, P. et al. (1992) Kidney Int. 41:220.
  6. Appay, V. and S.L. Rowland-Jones (2001) Trends Immunol. 22:83.
  7. Levy, J.A. (2009) J. Immunol. 182:3945.
  8. Randolph-Habecker, J.R. et al. (2002) Cytokine 19:37.
  9. DeVico, A.L. and Gallo, R.C. (2004) Nat. Rev. Microbiol. 2:401.
  10. Proost, P. et al. (1998) J. Biol. Chem. 273:7222.
  11. Appay, V. et al. (1999) J. Biol. Chem. 274:27505.
  12. Proudfoot, A.E.I. et al. (2003) Proc. Natl. Acad. Sci. USA 100:1885.
  13. von Hundelshausen, P. et al. (2005) Blood 105:924.
  14. von Hundelshausen, P. et al. (2001) Circulation 103:1772.
  15. Zernecke, A. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:1897.
  16. Baltus, T. et al. (2003) Blood 102:1985.
  17. Soria, G. and A. Ben-Baruch (2008) Cancer Lett. 267:271.

Alternate Names

RANTES, SISd

Entrez Gene IDs

6352 (Human); 20304 (Mouse); 403522 (Canine); 493689 (Feline)

Gene Symbol

CCL5

Additional CCL5/RANTES Products

Product Documents for Mouse CCL5/RANTES 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 Mouse CCL5/RANTES Antibody

For research use only

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