Human MCAF (MCP-1) Matched Antibody Pair for ELISpot

INTENDED USE

Pre-titered capture antibody and biotinylated detection antibody matched pair for the development of enzyme-linked immunospot (ELISpot) assays for the quantitation of single cells releasing human MCAF.

A recommended assay protocol is provided. The dilutions of capture antibody and detection antibody is determined according to this protocol. The researcher can optimize the dilutions if it is necessary.

For laboratory research use only. Not for use in diagnostic procedures. 

INTRODUCTION 

Monocyte chemotactic and activating factor (MCAF), also known as monocyte chemotactic protein 1 (MCP-1), lymphocyte–derived chemotactic factor (LDCF), and glioma-derived chemotactic factor (GDF), is a recently-identified chemotactic cytokine for monocytes.  CDNA cloning and structural analysis has revealed that this 76-amino acid polypeptide with a predicted molecular mass of 8,700 daltons belongs to a family of structurally-related low molecular weight proteins characterised by four conserved cysteine residues designated C-C family or intercrine b family (1,2,3). 

MCAF is expressed by various types of cultured cells including monocytes, lymphocytes, fibroblasts, endothelial cells, smooth muscle cells and transformed cell lines upon stimulation with LPS or cytokines such as IL-1, TNF-α, and IFN-γ.  Although there exist some minor differences in expression patterns that are observed in some types of cells, almost all agents that induce IL-8 mRNA expression also induce MCAF mRNA expression.  Platelet-derived growth factor is a strong inducer of MCAF mRNA in human fibroblasts whereas it failed to induce IL-8 mRNA in human fibroblasts (4).  These results suggest that the regulatory mechanism of MCAF gene expression differs from that of the IL-8 gene.  In addition to being chemotactic for monocytes, MCAF also activates human monocytes to become cytostatic for several human tumour cell lines (5), release lysosomal enzymes (6), and generate superoxide (6).

MCAF is also expressed in vivo by lung epithelial cells in patients with idiopathic pulmonary fibrosis (7), synovial tissues of rheumatoid arthritis (8), or in atheromatous plaques in atherosclerotic lesion (9), suggesting the participation of MCAF in the pathogenesis of these disorders.  Furthermore, MCAF has a potent histamine-releasing activity on basophils (10) that indicates an associated effect in allergic inflammations.  MCAF expression in vivo has been investigated by qualitative methods such as in situ hybridization and immunohistochemistry.  In order to further clarify and elucidate its participation and relation with various disorders, quantitative analysis of its in vivo level is necessary (11, 12).

This ELISpot kit is developed to detect and visualize of single cells secreting human MCAF.