Detects rat CCL4/MIP-1 beta in direct ELISAs and Western blots. In direct ELISAs, approximately 8% cross-reactivity with recombinant mouse CCL4/MIP-1 beta and recombinant rat CCL3/MIP-1 alpha is observed, and less than 2% cross-reactivity with recombinant human CCL4/MIP-1 beta is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Rat CCL4/MIP-1 beta Antibody

Detection of Rat CCL4/MIP‑1 beta 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 1 µg/mL of Sheep Anti-Rat CCL4/MIP-1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6935) 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 1.

Chemotaxis Induced by CCL4/MIP‑1 beta and Neutralization by Rat CCL4/MIP‑1 beta Antibody.

Recombinant rat CCL4/MIP-1 beta (Catalog # 6935-MB) induces BaF3 mouse proB 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 rat CCL4/MIP-1 beta (0.075 ug/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Rat CCL4/MIP-1 beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6935). The ND₅₀ is typically 0.1-0.5 ug/mL.

Applications for Rat CCL4/MIP-1 beta Antibody

Application

Recommended Usage

Western Blot

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

Neutralization

Measured by its ability to neutralize CCL4/MIP‑1 beta-induced chemotaxis in the BaF3 mouse proB cell line transfected with human CCR5. The Neutralization Dose (ND₅₀) is typically 0.1-0.5 ug/mL in the presence of 0.075 ug/mL Recombinant rat CCL4/MIP‑1 beta.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 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: 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 inflammation 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 down‑regulation of CCR5 (10). Mature rat CCL4 shares 80% and 86% aa sequence identity with human and mouse CCL4, respectively.

References

Rot, A. and U.H. von Andrian (2004) Annu. Rev. Immunol. 22:891.
Menten, P. et al. (2002) Cytokine Growth Factor Rev. 13:455.
Sun, X. et al. (2006) Infec. Immun. 74:5943.
Bisset, L.R. and Schmid-Grendelmeier, P. (2005) Curr. Opin. Pulm. Med. 11:35.
Frangogiannis, N.G. (2004) Inflamm. Res. 53:585.
Bystry, R.S. et al. (2001) Nat. Immunol. 2:1126.
Oliveira, S.H.P. et al. (2002) J. Leukoc. Biol. 71:1019.
Schall, T.J. et al. (1993) J. Exp. Med. 177:1821.
Guan, E. et al. (2001) J. Biol. Chem. 276:12404.
Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
Guan, E. et al. (2004) J. Cell. Biochem. 92:53.

Alternate Names

Exodus-3, MIP-1 beta, MIP1 beta

Entrez Gene IDs

6351 (Human); 20303 (Mouse); 116637 (Rat); 448786 (Canine); 102117861 (Cynomolgus Monkey)

Gene Symbol

CCL4

UniProt

P50230

Additional CCL4/MIP-1 beta Products

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Product Specific Notices for Rat CCL4/MIP-1 beta Antibody

For research use only

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Rat CCL4/MIP-1 beta Antibody

R&D Systems, part of Bio-Techne | Catalog # AF6935

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