Canine CCL4/MIP-1 beta Antibody

Catalog # Availability Size / Price Qty
AF5839
AF5839-SP
Detection of Canine CCL4/MIP‑1 beta  by Western Blot.
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Canine CCL4/MIP-1 beta Antibody Summary

Species Reactivity
Canine
Specificity
Detects canine CCL4/MIP‑1 beta in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human CCL4/MIP‑1 beta and recombinant mouse CCL4/MIP‑1 beta is observed and less than 1% cross-reactivity with recombinant viral CCL4/MIP‑1 beta is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant canine CCL4/MIP‑1 beta
Ala24-Asn92
Accession # NP_001005250
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
Immersion fixed canine peripheral blood mononuclear cells

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Example

Western Blot Detection of Canine CCL4/MIP‑1 beta  by Western Blot. View Larger

Detection of Canine CCL4/MIP‑1 beta by Western Blot. Western blot shows lysates of canine peripheral blood mononuclear cells (PBMC) untreated (-) or treated (+) with 5 µg/mL PMA and 500 ng/mL Ionomycin overnight. PVDF Membrane was probed with 1 µg/mL of Canine CCL4/MIP‑1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5839) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CCL4/MIP‑1 beta at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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: CCL4/MIP-1 beta

CCL4, also known as macrophage inflammatory protein 1 beta (MIP-1 beta, is a 12 kDa beta chemokine that is secreted at sites of inflammation by activated leukocytes, lymphocytes, vascular endothelial cells, and pulmonary smooth muscle cells (1, 2). CCL4 attracts a variety of immune cells to sites of microbial infection as well as to other pathologic inflammations such as allergic asthma and ischemic myocardium (3-8). A CCL4 deficiency in mice promotes the development of autoantibodies, possibly as a result of compromised regulatory T cell recruitment (6). CCL4 is secreted from activated monocytes as a heterodimer with CCL3/MIP-1 alpha (9). The first two N-terminal amino acids can be cleaved from human CCL4 by CD26/DPPIV (10, 11). Both the full length and truncated forms exert biological activity through CCR5, and the truncated form additionally interacts with CCR1 and CCR2 (10). In humans, the ability of CCL4 to bind CCR5 inhibits the cellular entry of M-tropic HIV-1 which utilizes CCR5 as a coreceptor (2). Both forms of CCL4 block HIV-1 infection of T cells by inducing the downregulation of CCR5 (10). Mature canine CCL4 shares 80-84% amino acid sequence identity with human, mouse, and rat CCL4.

References
  1. Rot, A. and U.H. von Andrian (2004) Annu. Rev. Immunol. 22:891.
  2. Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
  3. Sun, X. et al. (2006) Infec. Immun. 74:5943.
  4. Bisset, L.R. and P. Schmid-Grendelmeier (2005) Curr. Opin. Pulm. Med. 11:35.
  5. Frangogiannis, N.G. (2004) Inflamm. Res. 53:585.
  6. Bystry, R.S. et al. (2001) Nat. Immunol. 2:1126.
  7. Oliveira, S.H.P. et al. (2002) J. Leukoc. Biol. 71:1019.
  8. Schall, T.J. et al. (1993) J. Exp. Med. 177:1821.
  9. Guan, E. et al. (2001) J. Biol. Chem. 276:12404.
  10. Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
  11. Guan, E. et al. (2004) J. Cell. Biochem. 92:53.
Entrez Gene IDs
6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine)
Alternate Names
ACT2; ACT-2; AT744.1; C-C motif chemokine 4; CCL4; chemokine (C-C motif) ligand 4; Exodus-3; G-26 T-lymphocyte-secreted protein; G-26; HC21; LAG1; LAG-1; Lymphocyte activation gene 1 protein; Macrophage inflammatory protein 1-beta; MIP1 beta; MIP-1 beta; MIP1B; MIP1B1; MIP-1-beta; MIP-1-beta(1-69); PAT 744; Protein H400; SCYA2; SCYA4; secreted protein G-26; SIS-gamma; small inducible cytokine A4 (homologous to mouse Mip-1b); Small-inducible cytokine A4; T-cell activation protein 2

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