Human MCAF / MCP-1 ELISA Kit

RACTIVITY	Human
SENSITIVITY	<4 pg/mL
ASSAY RANGE	4-1600 pg/mL
REAGENTS PROVIDED	MCAF MICROTITER PLATE  MCAF CONJUGATE  MCAF STANDARD  CALIBRATOR DILUENT I  CALIBRATOR DILUENT II  WASH BUFFER (20X/96 wells, 30X/192 wells)  SUBSTRATE A  SUBSTRATE B  STOP SOLUTION

INTENDED USE

This Human MCAF ELISA kit is to be used for the in vitro quantitative determination of human monocyte chemotactic and activating factor (MCAF) concentrations in serum, plasma, cell culture supernatant, and other biological fluids.  This kit is intended for LABORATORY RESEARCH USE ONLY and is not to be used in diagnostic or therapeutic 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-a, and IFN-g.  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 MCAF ELISA is a 2.5-hour solid-phase immunoassay readily applicable to measure MCAF levels in serum, plasma, cell culture supernatant, and other biological fluids in the range of 0 to 1600 pg/mL.  It has shown no cross-reactivity with various other C-C and C-X-C chemokines IL-8 superfamily proteins.  This MCAF ELISA is expected to be effectively used for further investigation into the relationship between MCAF and various diseases.

CITATIONS

1. Elisa A. Liehn et al. A New Monocyte Chemotactic Protein-1/Chemokine CC Motif Ligand-2 Competitor Limiting Neointima Formation and Myocardial Ischemia/Reperfusion Injury in Mice. Journal of the American College of Cardiology Volume 56, Issue 22, 23 November 2010, Pages 1847–1857.

2. Margaret J. Burns and Martha B. Furie. Borrelia burgdorferi and Interleukin-1 Promote the Transendothelial Migration of Monocytes In Vitro by Different Mechanisms. Infect Immun. Oct 1998; 66(10): 4875–4883.

3. Sarchielli P et al. Chemokine levels in the jugular venous blood of migraine without aura patients during attacks.  Headache. 2004 Nov-Dec;44(10):961-8.

4. Kalayoglu MV et al.  Identification of Chlamydia pneumoniae within human choroidal neovascular membranes secondary to age-related macular degeneration. Graefes Arch Clin Exp Ophthalmol. 2005 Nov;243(11):1080-90. Epub 2005 May 21.

5. Boels MGS, Koudijs A, Avramut MC, Sol WMPJ, Wang G, van Oeveren-Rietdijk AM, van Zonneveld AJ, de Boer HC, van der Vlag J, van Kooten C, Eulberg D, van den Berg BM, IJpelaar DHT, Rabelink TJ. Systemic Monocyte Chemotactic Protein-1 Inhibition Modifies Renal Macrophages and Restores Glomerular Endothelial Glycocalyx and Barrier Function in Diabetic Nephropathy. Am J Pathol. 2017 Nov;187(11):2430-2440.

6. Chang M, Guo F, Zhou Z, Huang X, Yi L, Dou Y, Huan J. HBP induces the expression of monocyte chemoattractant protein-1 via the FAK/PI3K/AKT and p38 MAPK/NF-κB pathways in vascular endothelial cells. Cell Signal. 2018 Mar;43:85-94.