This immunoassay kit allows for the in vitro quantitative determination of mouse intercellular adhesion molecule 1,ICAM-1 concentrations in cell culture supernates, serum, plasma and other biological fluids.
Type d'échantillon
Cell Culture Supernatant, Plasma, Serum
Analytical Method
Quantitative
Specificité
This assay recognizes recombinant and natural mouse ICAM-1/CD54.
Réactivité croisée (Details)
No significant cross-reactivity or interference was observed.
Sensibilité
< 0.078 ng/mL The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest detectable concentration that could be differentiated from zero.
The microtiter plate provided in this kit has been pre-coated with an antibody specific to ICAM-1/CD54. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for ICAM-1/CD54 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain ICAM-1/CD54, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ICAM-1/CD54 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Restrictions
For Research Use only
Stock
4 °C/-20 °C
Stockage commentaire
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
Adhesion molecules mediate the interaction of cells with the extracellular matrix and with other cells. The immunoglobulin superfamily of proteins contains a large class of adhesion molecules with multiple immunoglobulin-like domains. ICAM is a member of this family. It is a 90 kDa type-I transmembrane glycoprotein with five Ig-like extracellular domains. The most important ligands for ICAM-1 are the _2 integrins LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18), which are expressed on leukocytes. ICAM-1 thus mediates the adhesion of leukocytes to ICAM-1-expressing cells. ICAM-1 also binds fibrinogen, hyaluronan, Rhinoviruses, Plasmodium falciparum-infected erythrocytes and CD43 (sialophorin) ICAM-1 is either a transmembrane protein (mICAM-1) or soluble (sICAM-1). mICAM-1 is expressed on endothelial and epithelial cells, lymphocytes, monocytes, eosinophils, keratinocytes, dendritic cells, hematopoietic stem cells, hepatocytes and fibroblasts. Regulation of ICAM-1 expression is cell specific. Up-regulation generally is by inflammatory cytokines (TNF- α , IFN- γ and IL-1) and down-regulation generally is by anti-inflammatory agents (e.g.glucocorticoids). One important, well-characterized function of ICAM-1 is immune-cell trafficking. At sites of inflammation, inflammatory cytokines induce up-regulation of ICAM-1 expression on vascular endothelial cells and activation of leukocyte integrins (LFA-1 and Mac-1). This leads to adhesion of leukocytes to the local endothelium, an essential step in migration of leukocytes to the site of inflammation. sICAM-1 has been reported in serum, cerebrospinal fluid and bronchoalveolar lavage. sICAM-1 likely arises by proteolytic cleavage of mICAM-1, synthesis from an alternatively spliced message has not been found. In general, elevated levels of serum ICAM-1 appear to be associated with inflammatory conditions and certain malignancies. It has, however, been pointed out that in inflammatory conditions, where the ligands LFA-1 and Mac-1 are likely to be activated, 2 binding and clearance of sICAM-1 might be enhanced, so that a reciprocal relationship between sICAM-1 levels and inflammation also is possible.