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Adhesion molecules array

Product Method Size Catalog Price
Adhesion molecules array B A T (evidence®) 4x45 (180 biochips) EV3572 POA
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Intended Use

Evidence Adhesion Molecules array is to be used for the in-vitro simultaneous quantitative detection of multiple related adhesion molecule immunoassays (in parallel) from a single sample.

This product is for research purposes only and not for diagnostic procedures.

Clinical Significance

Adhesion Molecules are complex membrane proteins that have an important role in a number of cellular processes. The Adhesion Molecules have been divided into 4 main families: the selectins, the immunoglobin (Ig) superfamily, integrins and cadherins.

In addition to the membrane forms, the adhesion molecules can be shed from the cell surface and can be detected in the blood of normal healthy persons with the levels being altered in a number of pathological states. Increased knowledge of these changes in the levels of the soluble adhesion molecules should help to further the understanding of their physiological role and pathological significance. This should lead to better diagnosis and disease management.

Principle

Evidence Biochip Array Technology is used to perform simultaneous quantitative detection of multiple analytes from a single sample. The core technology is the Randox Biochip, a 9 mm (2) solid substrate containing an array of discrete test regions of immobilized antibodies specific to different adhesion molecule markers. A chemiluminescent immunoassay is employed for the adhesion molecule assays. Increased levels of adhesion molecules in a specimen will lead to increased binding of antibody labeled with Horse Radish Peroxidase (HRP) and thus an increase in the chemiluminescence signal emitted.

The light signal generated from each of the test regions on the biochip is detected using digital imaging technology and compared to that from a stored calibration curve. From this the concentration is calculated.

Several different immunoassay based multi-analyte panels have been developed for use on Evidence. The adhesion molecules array will successfully quantitatively test for soluble E-selectin, L-selectin, P-selectin, ICAM-1 and VCAM-1 simultaneously.

 

Human Soluble E-Selectin (E-SEL) Assay

Intended Use

The Evidence human soluble E-selectin (E-SEL) assay has been designed for the quantitative measurement of hsE-selectin in human serum and plasma samples.

This assay is for research and development use only.

Clinical Significance

E-selectin is a member of the selectin family of adhesion molecules (1) . Selectins are transmembrane proteins which facilitate the establishment of the first relatively weak bonds between leukocytes and endothelial cells. The establishment of these bonds slows the flow of leukocytes across altered or injured endothelial cells. This process causes the activation of leukocyte integrins which allows firm adhesion of leukocytes on endothelial cells by interactions between the leukocyte integrins and members of the immunoglobin superfamily of adhesion molecules (2-3) .

E-selectin is a 70-115 kDa glycoprotein consisting of an extracellular aminoterminal calcium-dependent C2-type lectin domain, followed by an epidermal growth factor like domain, 6 short consensus repeats, a transmembrane domain and a short cytoplasmic tail. Adhesion by selectins is dependent on sialic acid and it has been suggested that fucose is also required. However it is thought that the selectins actually recognize a specific grouping of carbohydrate on particular glycoproteins (1) .

E-selectin is only expressed on endothelial cells and only after activation by inflammatory cytokines or endotoxin. Its expression is transitory and reaches a maximum 2 to 6 hours after cell activation (1,4) . It is then shed into the circulation where it may activate neutrophils and act as a pro-inflammatory agent (5) .

Soluble E-selectin can be detected in healthy persons and the levels have been shown to be altered in a number of pathological conditions (6-9) .

Principle

The Evidence E-SEL assay is a sandwich chemiluminescent assay for the detection of E-SEL in human serum and plasma.

REFERENCES

1. Kansas, G.S., Selectins and their ligands: current concepts and controversies. Blood. 1996; 88: 3259-3287.

2. Kyan-Aung, U., et al, Endothelial leukocyte adhesion molecule-1 and intercellular adhesion molecule-1 mediate the adhesion of eosinophils to endothelial cells in vitro and are expressed by endothelium in allergic cutaneous inflammation in vivo. J. Immunol. 1991; 146: 521-528.

3. Lawrence, M.B. and Springer, T.A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991; 65: 859-873.

4. Bevilacqua, M.P. et al, Endothelial leukocyte adhesion molecule 1: an inducible receptor for neutrophils related to complement regulatory proteins and lectins. Science. 1989; 243: 1160 -1165.

5. Ruchaud-Sparagano, M.H. et al, Potential pro-inflammatory effects of soluble E-selectin upon neutrophil function. Eur. J. Immunol. 1998; 28: 80-89.

6. Koch, A.E. et al, Soluble E-selectin in arthritis. Clin Immunol Immunopathol. 1993; 69: 29-35.

7. Cowley, H.C. et al, Increased circulating adhesion molecule concentrations in patients with the systemic inflammatory response syndrome: a prospective cohort study. Crit Care Med. 1994; 22: 651-657.

8. Banks, R.E. et al, Circulating intercellular adhesion molecule-1 (ICAM-1), E-selctin and vascular cell adhesion molecule-1 (VCAM-1) in human malignancies. Br. J. Cancer. 1993; 68: 122-124.

9. Fassbender, K. et al, Circulating selectin and immunoglobin-type adhesion molecules in acute ischemic stroke. Stroke 1995; 26: 1361-1364.

 

Human Soluble Intercellular Adhesion Molecule (ICAM-1) Assay

Intended Use

The Evidence ICAM-1 assay has been designed for the quantitative measurement of ICAM-1 in human serum and plasma samples.

This assay is for research and development use only.

Clinical Significance

ICAM-1 (CD54) is a member of the immunoglobin (Ig) like superfamily of adhesion molecules. The Ig like superfamily of adhesion molecules are transmembrane proteins which facilitate the establishment of the strong bonds between cells and the extracellular matrix.

ICAM-1 is a glycoprotein with five Ig-like extracellular domains, and a molecular weight of approximately 90 kDa. It is expressed on endothelial and epithelial cells as well as lymphocytes, monocytes, eosphinophils, kertanocytes, dendritic cells, hematopoietic stem cells, hepatocytes and fibroblasts. Its expression is generally transcriptionally regulated and cell specific. ICAM-1 plays an important role in inflammatory processes and in the T-cell mediated host defence system. Ligands of ICAM-1 include the membrane bound integrin receptors LFA-1 and Mac-1 on leukocytes, CD43, fibrinogen, hyaluronan and rhinoviruses (1) .

The membrane bound ICAM-1 is proteolytically cleaved and soluble ICAM-1 can be detected in serum from healthy persons (1-4) . It has also been measured in other body fluids (5-6) . It seems that all the soluble ICAM-1 arises from the shedding of the membrane bound ICAM-1. Altered levels of soluble ICAM-1 have been recorded in a number of pathological conditions (1-4, 7) .

Principle

The Evidence ICAM-1 assay is a sandwich chemiluminescent assay for the detection of ICAM-1 in human serum and plasma.

REFERENCES

1. Van de Stolpe, A. and van der Saag, P.T. Intercellular adhesion molecule-1. J Mol Med. 1996; 74: 13-33.

2. Gearing, A.J. et al, Soluble forms of vascular adhesion molecules, E-selectin, ICAM-1 and VCAM-1: pathological significance. Ann N Y Acad Sci. 1992; 667: 324-331.

3. Syrigos, K.N. et al, Prognostic significance of soluble adhesion molecules in Hodgkin's disease. Anticancer Res. 2004; 24: 1243-1247.

4. Zaccagni, H et al, Soluble adhesion molecule levels, neuropsychiatric lupus and lupus-related damage. Front Biosci. 2004; 9: 1654-1659.

5. Beeh, K.M. et al, Neutrophilic inflammation in induced sputum of patients with idiopathic pulmonary fibrosis. Sarcoidosis Vasc Diffuse Lung Dis. 2003; 20: 138-143.

6. Selakovic, V. et al, Cerebrospinal fluid and plasma concentration of soluble intercellular adhesion molecule 1, vascular cell adhesion molecule 1 and endothelial leukocyte adhesion molecule in patients with acute ischemic brain disease. Vojnosanit Pregl. 2003; 60: 139-146.

7. Guray, U et al, Levels of soluble adhesion molecules in various clinical presentations of coronary atherosclerosis. Int J Cardiol. 2004; 96: 235-240.

 

Human Soluble L-Selectin (L-SEL) Assay

Intended Use

The Evidence L-SEL assay has been designed for the quantitative measurement of hsL-selectin in human serum samples.

This assay is for research and development use only.

Clinical Significance

L-selectin is a member of the selectin family of adhesion molecules (1) . Selectins are transmembrane proteins which facilitate the establishment of the first relatively weak bonds between leukocytes and endothelial cells. The establishment of these bonds slows the flow of leukocytes across altered or injured endothelial cells. This process causes the activation of leukocyte integrins which allows firm adhesion of leukocytes to the endothelial cells by interactions between the leukocyte integrins and members of the immunoglobin superfamily of adhesion molecules (2-3) .

L-selectin is a cell surface glycoprotein consisting of an extracellular amino terminal calcium-dependent C2-type lectin domain, followed by an epidermal growth factor like domain, 2 short consensus repeats, a transmembrane domain and a short cytoplasmic tail (1) . Unlike the other members of the selectin family L-selectin is not expressed on endothelial cells but has only been found on leukocytes. There are two forms of L-selectin depending on the cell type. The molecular weight of the lymphocyte form is approximately 74 kDa and the neutrophil form is approximately 100 kDa (4) .

Adhesion by selectins is dependent on sialic acid and it has been suggested that fucose is also required. However it is thought that the selectins actually recognize a specific grouping of carbohydrate on particular glycoproteins. A number of different ligands for L-selectin on endothelial cells have been identified and described (1) .

The L-selectin is shed from the cell surface by proteolytic degradation and this seems to be the main source of soluble L-selectin. The soluble L-selectin is slightly smaller than the membrane bound molecule and is biologically active in circulation. High levels can prevent the binding of lymphocytes to the endothelium (5) .

Soluble L-selectin can be detected in healthy persons and the levels have been shown to be altered in a number of pathological conditions (5-7) .

Principle

The Evidence L-SEL assay is a sandwich chemiluminescent assay for the detection of L-SEL in human serum and plasma.

REFERENCES

1. Kansas, G.S., Selectins and their ligands: current concepts and controversies. Blood. 1996; 88:3259-3287.

2. Kyan-Aung, U., et al, Endothelial leukocyte adhesion molecule-1 and intercellular adhesion molecule-1 mediate the adhesion of eosinophils to endothelial cells in vitro and are expressed by endothelium in allergic cutaneous inflammation in vivo. J. Immunol. 1991; 146: 521-528.

3. Lawrence, M.B. and Springer, T.A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991; 65: 859-873.

4. Lewinsohn, D.M. et al, Leukocyte-endothelial cell recognition: evidence of a common molecular mechanism shared by neutrophils, lymphocytes, and other leukocytes. J. Immunol. 1987; 138: 4313-4321.

5. Schleiffenbaum, B.E. et al, Soluble L-selectin is present in human plasma at high levels and retains functional activity. J Cell Biol. 1992; 119: 229-238.

6. Spertini, O. et al, ELISA for quantification of L-selectin shed from leukocytes in vivo. J Immunol Methods 1992; 156: 115-123.

7. Ates, A. et al, Serum-soluble selectin levels in patients with rheumatoid arthritis and systemic sclerosis. Scand J Immunol. 2004; 59: 315-320.

Kourtis, A.P. et al, Soluble L-selectin, a marker of immune activation, in neonatal infection. Clin Immunol. 2003; 109: 224-228.

 

Human Soluble P-Selectin (P-SEL) Assay

Intended Use

The Evidence P-SEL assay has been designed for the quantitative measurement of hsP -selectin in human serum and plasma samples.

This assay is for research and development use only.

Clinical Significance

P-selectin is a member of the selectin family of adhesion molecules (1) . Selectins are transmembrane proteins which facilitate the establishment of the first relatively weak bonds between leukocytes and endothelial cells. The establishment of these bonds slows the flow of leukocytes across altered or injured endothelial cells. This process causes the activation of leukocyte integrins which allows firm adhesion of leukocytes on endothelial cells by interactions between the leukocyte integrins and members of the immunoglobin superfamily of adhesion molecules (2-3) .

P-selectin is a cell surface glycoprotein consisting of an extracellular aminoterminal calcium-dependent C2-type lectin domain, followed by an epidermal growth factor like domain, 9 short consensus repeats, a transmembrane domain and a short cytoplasmic tail. It is 140 kDa and highly glycosylated (1) . Adhesion by selectins is dependent on sialic acid and it has been suggested that fucose is also required. However it is thought that the selectins actually recognize a specific grouping of carbohydrate on particular glycoproteins. P-selectin binds specifically to a glycoprotein termed P-selectin glycoprotein ligand-1 (PSGL-1) found on myeloid cells, blood neutrophils, monocytes and lymphocytes (1) .

P-selectin is associated with the α-granules in resting platelets and is rapidly redistributed to the cell surface upon activation. It is also found in a pre-formed state in the Weibel-Palade bodies of endothelial cells and similar to the platelets it is rapidly moved to the cell surface following stimulation (4) . After expression on the cell surface P-selectin is rapidly internalised and recycled or degraded within the cell. Stimulation by inflammatory mediators are also able to activate transcription of P-selectin mRNA (5) .

Soluble P-selectin is smaller than the membrane bound P-selectin and may be a proteolytic fragment, most being derived from the platelets rather than the endothelium. It could be a soluble variant lacking the transmembrane domain as an alternatively spliced mRNA encoding a form of P-selectin has been identified (6-9) .

Soluble P-selectin can be detected in healthy persons and the levels have been shown to be altered in a number of pathological conditions (9-11) .

Principle

The Evidence P-SEL assay is a sandwich chemiluminescent assay for the detection of P-SEL in human serum and plasma.

REFERENCES

1. Kansas, G.S., Selectins and their ligands: current concepts and controversies. Blood. 1996; 88: 3259-3287.

2. Kyan-Aung, U., et al, Endothelial leukocyte adhesion molecule-1 and intercellular adhesion molecule-1 mediate the adhesion of eosinophils to endothelial cells in vitro and are expressed by endothelium in allergic cutaneous inflammation in vivo. J. Immunol. 1991; 146: 521-528.

3. Lawrence, M.B. and Springer, T.A. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell. 1991; 65: 859-873.

4. Tedder, T.F. et al, The selectins: vascular adhesion molecules. FASEB J. 1995; 9: 866-873.

5. Weller, A. et al, Cloning of the mouse endothelial selectins. J. Biol. Chem. 1992; 267: 15176-15183.

6. Lasky, L.A. Selectin-carbohydrate interactions and the initiation of the inflammatory response. Annu Rev Biochem. 1995; 64: 113-139.

7. Dunlop, L.C. et al, Characterization of GMP-140 (P-selectin) as a circulating plasma protein. J Exp Med. 1992; 175: 1147-1150.

8. Ushiyama, S. et al, Structural and functional characterization of monomeric soluble P-selectin and comparison with membrane P-selectin. J. Biol. Chem. 1993; 268: 15229-15237

9. Mukae, H. et al, Elevated levels of circulating adhesion molecules in patients with active pulmonary tuberculosis. Respirology 2003; 8: 326-331.

10. Blann, A.D. et al, The adhesion molecule P-selectin and cardiovascular disease. Eur Heart J. 2003; 24: 2166-2179.

11. Ferroni, P. et al, Prognostic value of soluble P-selectin levels in colorectal cancer. Int J Cancer 2004; 111: 404-408.


Human Soluble Vascular Adhesion Molecule (VCAM-1) Assay

Intended Use

The Evidence VCAM-1 assay has been designed for the quantitative measurement of hsVCAM-1 in human serum and plasma samples.

This assay is for research and development use only.

Clinical Significance

VCAM-1 is a member of the immunoglobin (Ig) like superfamily of adhesion molecules. The Ig like superfamily of adhesion molecules are transmembrane proteins which facilitate the establishment of the strong bonds between leukocytes and the altered or activated endothelial cells. These interactions are critical for leukocyte-transendothelial migration during immune responses.

VCAM-1 is a glycoprotein with seven Ig-like extracellular domains, and a molecular weight of approximately100-110 kDa (1) .

As well as being expressed on endothelial cells, VCAM-1 is expressed on smooth muscle cells, fibroblasts, activated neurons, dendritic cells and macrophages (2-6) . The main receptor for VCAM-1 is α 4 β 1 /VLA-4 with it also binding to α 4 β 7 /LPAM-1. The VLA-4 receptor is expressed on all leukocytes except neutrophils (7) .

Principle

The Evidence VCAM-1 assay is a sandwich chemiluminescent assay for the detection of VCAM-1 in human serum and plasma.

REFERENCES

1. van de Stolpe, A. and van der Saag, P.T. Intercellular adhesion molecule-1. J Mol Med. 1996; 74: 13-33.

2. Gearing, A.J. et al, Soluble forms of vascular adhesion molecules, E-selectin, ICAM-1 and VCAM-1: pathological significance. Ann N Y Acad Sci. 1992; 667: 324-331.

3. Syrigos, K.N. et al, Prognostic significance of soluble adhesion molecules in Hodgkin's disease. Anticancer Res. 2004; 24: 1243-1247.

4. Zaccagni, H et al, Soluble adhesion molecule levels, neuropsychiatric lupus and lupus-related damage. Front Biosci. 2004; 9: 1654-1659.

5. Beeh, K.M. et al, Neutrophilic inflammation in induced sputum of patients with idiopathic pulmonary fibrosis. Sarcoidosis Vasc Diffuse Lung Dis. 2003; 20: 138-143.

6. Selakovic, V. et al, Cerebrospinal fluid and plasma concentration of soluble intercellular adhesion molecule 1, vascular cell adhesion molecule 1 and endothelial leukocyte adhesion molecule in patients with acute ischemic brain disease. Vojnosanit Pregl. 2003; 60: 139-146.

7. Guray, U et al, Levels of soluble adhesion molecules in various clinical presentations of coronary atherosclerosis. Int J Cardiol. 2004; 96: 235-240.