Name
sMAdCAM-1, Human, ELISA kit
Catalog nr
HK337
(lot number and expiry date are indicated on the label)
Short description
The mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a cell-surface Ig superfamily member composed of two extracellular Ig domains, followed by a mucin-like domain, a transmembrane domain and a short cytoplasmatic domain. It interacts via its N-terminal Ig domain with the lymphocyte homing receptor integrin alpha4beta7. MAdCAM-1 promotes the adhesion of T and B cells, monocytes/macrop...
Size
2 x 96 det.
Application
Manual
Description
The mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a cell-surface Ig superfamily member composed of two extracellular Ig domains, followed by a mucin-like domain, a transmembrane domain and a short cytoplasmatic domain. It interacts via its N-terminal Ig domain with the lymphocyte homing receptor integrin alpha4beta7. MAdCAM-1 promotes the adhesion of T and B cells, monocytes/macrophages, and potentially eosinophils, basophils, and differentiated mast cells to the vascular endothelium and is critical for lymphocyte homing to the gut. RNA transcripts are predominantly expressed in the small intestine, mesenteric lymph nodes, colon and spleen. MAdCAM-1 transcripts are weakly expressed in human pancreas and brain. The MAdCAM-1 protein (60 kDa) is widely expressed on endothelia in both lymphoid and non-lymphoid tissues. Furthermore, it is expressed in thymic medulla and on high endothelial venules (HEV). This expression increases upon response to several cytokines including TNF-alpha, IL1b and IFN-gamma. MAdCAM-1 expression is upregulated on HEV-like vessels in a variety of chronic inflammatory diseases, and may mediate increased leukocyte trafficking into inflamed tissue.
In utero and during early childhood, MAdCAM-1 plays a dominant role in lymphocyte-endothelial cell adhesion at both mucosal and nonmucosal sites. In contrast, in the adult the expression of MAdCAM-1 is restricted to mucosal tissues and has been shown to be dramatically up-regulated during intestinal inflammation. In the gut, MADCAM-1 is basically expressed on follicular dendrites in Peyer's patches. Its expression is dramatically increased in inflammatory bowel disease (IBD). It is expressed in animal models of IBD and in human tissue samples from patients with Crohn's disease and ulcerative colitis. MAdCAM-1 is strongly expressed in the synovium of osteoarthritis patients, predominantly on the endothelial lining of blood vessels, but also within the vessel lumen. The MAdCAM-1/integrin alpha4beta7 homing system possibly participates in gastric inflammation in response to Helicobacter pylori infection and contributes to mucosa-associated lymphoid tissue (MALT) formation typically leading to the development of nodular gastritis.
Higher expression of MAdCAM-1 is reflected in elevated levels of the circulating soluble form of MAdCAM-1 (sMAdCAM-1). Since MAdCAM-1 is elevated in inflammatory, infectious and malignant diseases, sMAdCAM-1 serves as a perfect non-invasive biomarker for disease acitivity. In sera of healthy donor, sMAdCAM-1 was detected at 236.5 ± 55.8 ng/ml. In urine of healthy donors, sMAdCAM-1 was detected at 20-123 ng/ml. Measurement of sMAdCAM-1 levels is potentially useful to monitor disease activity and the results of therapy.
Application
The human sMAdCAM-1 ELISA has been developed for the quantitative measurement of natural and recombinant sMAdCAM-1 in serum, plasma, urine, cell culture medium and breast milk. In serum or plasma samples, human sMAdCAM-1 can be measured accurately if serum or plasma samples are diluted at least 10 times. Most reliable results are obtained if EDTA plasma is used. Urine samples have to be diluted at least 5 times.
Features
- Minimum concentration which can be measured is 0.4 ng/ml sMAdCAM-1.
- Measurable concentration range of 0.4-100 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human sMAdCAM-1 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours.
- The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay.
- Samples and standards are captured by a solid bound specific antibody.
- Biotinylated tracer antibody will bind to captured sMAdCAM-1.
- Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
- Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
- The enzyme reaction is stopped by the addition of citric acid.
- The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human sMAdCAM-1 standards (log).
- The human sMAdCAM-1 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents, except for the standard, are stable for 1 month if stored at 2-8°C.
After reconstitution the standard is stable for 24 hours. For longer stability we recommend to store aliquots at
-20°C.
Recovery
Normal human blood samples (plasma), containing baseline levels of 10 ng/ml sMAdCAM-1, were spiked with sMAdCAM-1 in concentrations of 10 and 25 ng/ml. Samples with and without sMAdCAM-1 were incubated for 1 hour at room temperature. Samples were measured using the ELISA. Values for sMAdCAM-1 ranged between 67 % and 122 % (mean 93%).
Linearity
The linearity of the assay was determined by serially diluting a sample containing 50 ng/ml human sMAdCAM-1. The diluted samples were measured in the assay. The line obtained a slope of 0.96 and a correlation coefficient of 0.994. 
References
- Leung, E et al; Bioassay detects soluble MAdCAM-1 in body fluids. Immunol Cell Biol 2004, 82: 400
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use or derivation of this product.
Also available
References
- Leung, E et al; Bioassay detects soluble MAdCAM-1 in body fluids. Immunol Cell Biol 2004, 82: 400
Scientific info
sMAdCAM-1 as biomarker for inflammatory activity
Mucosal Addressin Cell Adhesion Molecule-1
The mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a cell-surface Ig superfamily member of ~60 kDa. Although MAdCAM-1 is preferentially expressed on the surface of high endothelial venules (HEV) in the gut and associated lymphoid tissue (Peyer’s patches), it can also be detected in the mammary gland during lactation and on brain and hepatic endothelial cells.
MAdCAM-1 and lymphocyte interaction
MAdCAM-1 promotes the adhesion of T and B cells, monocytes/macrophages, and potentially eosinophils, basophils, and differentiated mast cells to the vascular endothelium and is critical for lymphocyte homing to the gut. In inflammatory bowel disease (IBD), especially Crohn’s disease, MAdCAM-1 acts as the main ligand for alpha4beta7-expressing lymphocytes, and recruits these lymphocytes into the intestine where they can initiate and sustain chronic inflammation.
MAdCAM-1 in inflamed tissue
MAdCAM-1 upregulation has been detected on HEV in chronically inflamed pancreas, on chronically inflamed pancreas of the non-obese diabetic mouse, on endothelia and choroid plexus epithelial cells in chronic relapsing experimental autoimmune encephalomyelitis (EAE), and on gut lamina propria venules in IBD. Since MAdCAM-1 was found to be upregulated on hepatic endothelial cells in response to cytokines, it is presumably also involved in chronic liver inflammation.
Soluble MAdCAM-1 as inflammatory marker
Higher expression of MAdCAM-1 is reflected in elevated levels of the circulating soluble form of MAdCAM-1 (sMAdCAM-1). Since MAdCAM-1 is elevated in inflammatory, infectious and malignant diseases, sMAdCAM-1 serves as a perfect non-invasive biomarker for disease activity. Monitoring the presence and levels of sMAdCAM-1 in body fluids may be helpful to diagnose inflammation in humans. In sera of healthy donor, sMAdCAM-1 was detected at 236.5 ± 55.8 ng/ml. In urine of healthy donors, sMAdCAM-1 was detected at 20-123 ng/ml.
In conclusion, the Hbt Human sMAdCAM-1 ELISA is a sensitive, non-invasive tool for monitoring chronic inflammatory disease activity in a diverse field of inflammatory research.
Special features:
• Useful for quantitative measurement of human sMAdCAM-1 in plasma, urine, mother milk and other body fluids
• Standard curve: 0.4 to 100 ng/ml
• Sample volume: less than 25 ul plasma per determination
Selected reading
1. Ando T et al; Inflammatory cytokines induce MAdCAM-1 in murine hepatic endothelial cells and mediate alpha-4 beta-7 integrin dependent lymphocyte endothelial adhesion. BMC Physiology 2007, 7: 10
2. Bachmann C et al; Targeting mucosal addressin cellular adhesion molecule (MAdCAM)-1 to noninvasively image experimental Crohn’s disease. Gastroenterol 2006, 130: 8
3. Connor E et al; Expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in acute and chronic inflammation. J Leukoc Biol 1999, 65: 349
4. Farkas S et al; Blocking MAdCAM-1 in vivo reduces leukocyte extravasation and reverses chronic inflammation in experimental colitis. Int J Colorectal Dis 2006, 21: 71
5. Leung E et al; Bioassay detects soluble MAdCAM-1 in body fluids. Immunol Cell Biol 2004, 82: 400
6. Xu Y et al; The correlation between proinflammatory cytokines, MAdCAM-1 and cellular infiltration in the inflamed colon from TNF-alpha gene knockout mice. Immunol Cell Biol 2007, 85: 633
Mucosal Addressin Cell Adhesion Molecule-1
The mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a cell-surface Ig superfamily member of ~60 kDa. Although MAdCAM-1 is preferentially expressed on the surface of high endothelial venules (HEV) in the gut and associated lymphoid tissue (Peyer’s patches), it can also be detected in the mammary gland during lactation and on brain and hepatic endothelial cells.
MAdCAM-1 and lymphocyte interaction
MAdCAM-1 promotes the adhesion of T and B cells, monocytes/macrophages, and potentially eosinophils, basophils, and differentiated mast cells to the vascular endothelium and is critical for lymphocyte homing to the gut. In inflammatory bowel disease (IBD), especially Crohn’s disease, MAdCAM-1 acts as the main ligand for alpha4beta7-expressing lymphocytes, and recruits these lymphocytes into the intestine where they can initiate and sustain chronic inflammation.
MAdCAM-1 in inflamed tissue
MAdCAM-1 upregulation has been detected on HEV in chronically inflamed pancreas, on chronically inflamed pancreas of the non-obese diabetic mouse, on endothelia and choroid plexus epithelial cells in chronic relapsing experimental autoimmune encephalomyelitis (EAE), and on gut lamina propria venules in IBD. Since MAdCAM-1 was found to be upregulated on hepatic endothelial cells in response to cytokines, it is presumably also involved in chronic liver inflammation.
Soluble MAdCAM-1 as inflammatory marker
Higher expression of MAdCAM-1 is reflected in elevated levels of the circulating soluble form of MAdCAM-1 (sMAdCAM-1). Since MAdCAM-1 is elevated in inflammatory, infectious and malignant diseases, sMAdCAM-1 serves as a perfect non-invasive biomarker for disease activity. Monitoring the presence and levels of sMAdCAM-1 in body fluids may be helpful to diagnose inflammation in humans. In sera of healthy donor, sMAdCAM-1 was detected at 236.5 ± 55.8 ng/ml. In urine of healthy donors, sMAdCAM-1 was detected at 20-123 ng/ml.
In conclusion, the Hbt Human sMAdCAM-1 ELISA is a sensitive, non-invasive tool for monitoring chronic inflammatory disease activity in a diverse field of inflammatory research.
Special features:
• Useful for quantitative measurement of human sMAdCAM-1 in plasma, urine, mother milk and other body fluids
• Standard curve: 0.4 to 100 ng/ml
• Sample volume: less than 25 ul plasma per determination
Selected reading
1. Ando T et al; Inflammatory cytokines induce MAdCAM-1 in murine hepatic endothelial cells and mediate alpha-4 beta-7 integrin dependent lymphocyte endothelial adhesion. BMC Physiology 2007, 7: 10
2. Bachmann C et al; Targeting mucosal addressin cellular adhesion molecule (MAdCAM)-1 to noninvasively image experimental Crohn’s disease. Gastroenterol 2006, 130: 8
3. Connor E et al; Expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) in acute and chronic inflammation. J Leukoc Biol 1999, 65: 349
4. Farkas S et al; Blocking MAdCAM-1 in vivo reduces leukocyte extravasation and reverses chronic inflammation in experimental colitis. Int J Colorectal Dis 2006, 21: 71
5. Leung E et al; Bioassay detects soluble MAdCAM-1 in body fluids. Immunol Cell Biol 2004, 82: 400
6. Xu Y et al; The correlation between proinflammatory cytokines, MAdCAM-1 and cellular infiltration in the inflamed colon from TNF-alpha gene knockout mice. Immunol Cell Biol 2007, 85: 633
MSDS
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