Soluble Epoxide Hydrolase Cell-Based Assay Kit Reference: 600090-480 Mammalian sEH is a member of the α/β-hydrolase fold family of enzymes that catalyze the hydrolysis of exogenous and endogenous epoxides to vicinal diols. Endogenous substrates for sEH include EETs which exhibit vasodilatory and anti-inflammatory activity. Cayman’s sEH Cell-Based Assay provides a convenient 96-well plate, fluorescence-based method for detecting epoxide hydrolase activity in whole cells. The assay utilizes Epoxy Fluor 7, a sensitive fluorescent substrate for sEH that can be used to monitor the activity of both human and murine enzymes.
7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit Reference: 600120-1 Cayman’s 7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit employs CFSE to label target cells within the mixed cell population and 7-AAD to label dead cells. This kit provides an improvement over the traditional 51chromium (51Cr) release assay to assess cell-mediated cytotoxicity. CFSE labeling is more sensitive, does not employ radioisotopes, and cytolysis can be assessed at the single-cell level. The mixed lymphocyte reaction is a cornerstone method of immunology(17323), and this kit can help immunologists to detect subtle changes in cytotoxic lymphocyte function. The kit provides sufficient reagents to effectively stain approximately 109 cells.
Autophagy/Cytotoxicity Dual Staining Kit Reference: 600140-1 Cayman’s Autophagy/Cytotoxicity Dual Staining provides a convenient tool for studying the regulation of autophagy and cytotoxicity at the cellular level. The kit employs MDC, an autofluorescent substance incorporated into multilamellar bodies by both an ion trapping mechanism and the interaction with membrane lipids, as a probe for detection of autophagic vacuoles in cultured cells. Propidium iodide is used as a marker of cell death.
Methionine Sulfoxide Immunoblotting Kit Reference: 600160-10 Protein MetO is a reversible oxidative modification that occurs by exposure of protein(s) methionine residues to reactive oxygen species (ROS). Methionine oxidation can alter the function(s) of the modified proteins and if not reversed by MetO reductases can be further oxidized to methionine sulfone, an irreversible modification. Cayman’s MetO Immunoblotting Kit contains reagents needed for the immunochemical detection of proteins containing MetO residues by western blotting. MetO-containing samples of interest include those from cell or tissue lysates as well as semi-pure or purified proteins.
Melanocortin-3 Receptor Reporter Assay Kit Reference: 600180-100 Cayman's MC3R Reporter Assay consists of a 96-well plate coated with a transfection complex containing DNA constructs for MC3R and a cAMP response element regulated Secreted Alkaline Phosphatase (SEAP) reporter (MC3R Reverse Transfection Strip Plate). Cells grown on the transfection complex will express MC3R at the cell surface. Binding of agonists to MC3R initiates a signal transduction resulting in expression of SEAP. SEAP activity is measured following addition of luminescence-based alkaline phosphatase substrate provided in the kit.
Melanocortin-4 Receptor Reporter Assay Kit Reference: 600190-100 Cayman’s MC4R Reporter Assay consist of a 96-well plate coated with a transfection complex containing DNA constructs for MC4R and a cAMP response element regulated Secreted Alkaline Phosphatase (SEAP) reporter (MC4R Reverse Transfection Strip Plate). Cells grown on the transfection complex will express MC4R at the cell surface. Binding of agonists to MC4R initiates a signal transduction cascade resulting in expression of SEAP which is secreted into the cell culture media. SEAP activity is measured following addition of a luminescence-based alkaline phosphatase substrate provided in the kit.
Orexin 1 Receptor Reporter Assay Kit Reference: 600240-100 Cayman's OX1R Reporter Assay Kit consists of a 96-well plate coated with a transfection complex containing DNA constructs for OX1R, a recombinant G-protein, and a cAMP response element regulated Secreted Alkaline Phosphatase (SEAP) reporter (OX1R Reverse Transfection Strip Plate). Cells grown on the transfection complex will express OX1R at the cell surface and the recombinant G-protein inside the cell. Binding of agonists to OX1R initiates a signal transduction cascade resulting in expression of SEAP which is secreted into the cell culture medium. Aliquots of culture medium are removed at time intervals beginning at about 16 hours and SEAP activity is measured following addition of a luminescence-based alkaline phosphatase substrate provided in the kit. The kit is easy to use and can be easily adapted to high throughput screening for therapeutic compounds regulating activation of OX1R. A known OX1R agonist, Orexin A, is included in the kit for use as a positive control. The kit provides sufficient reagent to measure SEAP activity at three time points, using the black plates provided.
Orexin 2 Receptor Reporter Assay Kit Reference: 600250-100 Cayman's OX2R Reporter Assay Kit consists of a 96-well plate coated with both OX2R and Secreted Alkaline Phosphatase (SEAP) reporter constructs (OX2R Reverse Transfection Strip Plate). Cells grown on the transfection complex will express OX2R at the cell surface and the recombinant G-protein inside the cell. Binding of agonists to OX2R initiates a signal transduction cascade resulting in expression of SEAP which is secreted into the cell culture medium. Aliquots of culture medium are removed at time intervals beginning at about 16 hours and SEAP activity is measured following addition of a luminescence-based alkaline phosphatases substrate provided in the kit. The kit is easy to use and can be easily adapted to high throughput screening for therapeutic compounds regulating activation of OX2R. A known OX2R agonist, Orexin A, is included in the kit for use as a positive control. The kit provides sufficient reagent to measure SEAP activity at three time points, using the black plates provided.
Secreted Alkaline Phosphatase Reporter Gene Assay Kit (Luminescence) Reference: 600260-300 SEAP is commonly used as a reporter of gene expression. Compared to other conventional intracellular reporters such as CAT and firefly luciferase, SEAP has the advantage of being secreted from transfected cells into the culture medium. Cayman’s Secreted Alkaline Phosphatase Reporter Gene Assay Kit (Luminescence) provides a simple chemiluminescence method for the sensitive quantitation of SEAP in conditioned cell culture medium from transfected cells.
Annexin V FITC Assay Kit Reference: 600300-100 One of the hallmarks of the early stages of apoptosis is the redistribution of membrane phospholipids such as phosphatidylserine and phosphatidylethanolamine from the inner to outer leaflet of the membrane bilayer where they are exposed on the cell surface. Externalization of phosphatidylserine residues to the outer plasma membrane leaflet allows their detection via their high affinity for annexin V, a phospholipid binding protein. Cayman’s Annexin V Assay Kit employs FITC-conjugated Annexin V as a probe for phosphatidylserine on the outer membrane of apoptotic cells.
Sulfenylated Protein Cell-Based Detection Kit Reference: 600320-96 Reactive oxygen species (ROS) react with proteins, resulting in protein modification, such as protein sulfenylation through the reversible oxidation of cysteine residues. Redox-sensitive cysteine residues in proteins can function as sensors of ROS and serve as molecular switches, activating or deactivating proteins, following a change in oxidative state. However, the accumulation of proteins with irreversible cysteine oxidation is a hallmark of stress-induced cellular damage associated with diseases like cancer. Cayman’s Sulfenylated Protein Cell-Based Detection Kit employs the cell-permeable and chemoselective DAz-2 probe to detect sulfenic acid-modified proteins in living cells. It can be used to monitor intracellular sulfenylated protein levels in living cells to discriminate between normal and pathological conditions or to help identify new pathways regulated by sulfenic acid formation.
EP2 Receptor (rat) Reporter Assay Kit Reference: 600340-100 The EP2 receptor is one of four GPCRs that mediate the actions of PGE2. The diverse effects of PGE2 acting via EP2 receptors point to the need to identify novel agonists and antagonists, both to further elucidate the function of this receptor subtype and for use as therapeutics for various diseases. Cayman's EP2 Receptor (rat) Reporter Assay Kit consists of a 96-well plate coated with a transfection complex containing DNA constructs for rat EP2 receptor and a cAMP response element regulated Secreted Alkaline Phosphatase (SEAP) reporter (EP2 Receptor Reverse Transfection Plate). Cells grown on the transfection complex will express EP2 at the cell surface. Binding of agonists to EP2 initiates a signal transduction cascade resulting in expression of SEAP which is secreted into the cell culture media. Aliquots of media are removed at time intervals, beginning at approximately six hours, and SEAP activity is measured following addition of a luminescence-based alkaline phosphatase substrate provided in the kit. The kit is simple to use and can be easily adapted to high throughput screening for therapeutic compounds regulating the activation of EP2. PGE2 is included in the kit for use as a positive control. The kit provides sufficient reagents to measure SEAP activity at three time points using the black plates included.