Rat CCL4/MIP-1 beta Antibody

Catalog #: AF6935 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6935
AF6935-SP

Detection of Rat CCL4/MIP‑1 beta by Western Blot.

2 Images

Product Details

Supplemental Products

Reviews

Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Rat CCL4/MIP-1 beta Antibody Summary

Species Reactivity

Rat

Specificity

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.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant rat CCL4/MIP-1 beta
Ala24-Asn92
Accession # P50230

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.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

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.

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.

Scientific Data

Western Blot

View Larger

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.

Neutralization

View Larger

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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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.

Entrez Gene IDs

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

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