Monoclonal antibody MNA.1 (formerly known as 5D3-F7) recognizes human natural and recombinant monocyte chemotactic protein-1 (MCP-1). Monocyte chemotactic protein-1 (MCP-1) is a 11 kDa protein belonging to the CC subgroup of the chemokine superfamily, which stimulate the migration of monocytic cells. In contrast, the CXC chemokines predominantly activate polymorphonuclear leukocytes. The coordinated synthesis and release of MCP-1 plays a central role in both acute and chronic inflammatory processes by controlling the influx of phagocytic cells. Furthermore, their state of activation is in concert with primary inflammatory cytokines, such as IL-1, TNF-a, and IL-6. A selective accumulation of MCP-1 in the cerebrospinal fluid (CSF) of AIDS patients with cytomegalovirus encephalitis, but not with other opportunistic infections or primary lymphomas of the central nervous system , has been described. Furthermore, the chemotactic activity of MCP-1 on monocytic cells has been suggested to play a role in psoriasis, rheumatoid arthritis and atherosclerosis. No cross-reactivity of mAb MNA.1 with other cytokines has been detected.
Flow cytometry, Frozen sections, Functional studies, Immuno assays, Immuno precipitation, Paraffin sections, Western blot
FC: Antibody MNA.1 stains intracellular MCP-1.For intracellular staining HUVEC cells were permeabilized with buffer containing- 0.1% saponin. The cells were fixed in 4% paraformaldehyde before staining. As negative control a corresponding isotype control antibody was used- (Ref.5).
W: A reduced sample treatment and 15% SDS-Page was used. The band sizes are ~14 and 11 kDa (Ref.1).
IHC-P: Tissue sections were fixed in formalin and pretreated with hydrogen peroxide to quench endogenous peroxidases. Antigen retrieval was performed by pressure cooking for 3 minutes in PBS. As negative control the MNA.1 antibody was omitted (Ref.7).
IHC-F:- Tissue sections were air dried and pretreated with 3% hydrogen peroxide to quench endogenous peroxidases. As negative control the MNA.1 antibody was omitted (Ref.2).
FS: Antibody MNA.1 inhibits migration of monocytes by neutralizing MCP-1(Ref.1).
For immunohistochemistry, flow cytometry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies- dilutions have to be optimized in user’s experimental setting.