PPARα, δ, γ Complete Transcription Factor Assay Kit Reference: 10008878-96 PPARs are ligand-activated transcription factors belonging to the large superfamily of nuclear receptors. PPARα primarily activates genes encoding proteins involved in fatty acid oxidation, while PPARγ primarily activates genes directly involved in lipogenic pathway and insulin signaling. Members of the PPAR family are important direct targets of many antidiabetic and hypolipidemic drugs. Cayman’s PPAR Transcription Factor Assays are a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysates.
WST-1 Cell Proliferation Assay Kit Reference: 10008883-96 Cayman’s WST-1 Proliferation Assay is based on the reduction of tetrazolium salt WST-1 to a soluble formazan dye by electron transport across the plasma membrane of dividing cells. This kit will also allow investigators to screen drug candidates involved in cell cycle regulation.
WST-1 Cell Proliferation Assay Kit Reference: 10008883-480 Cayman’s WST-1 Proliferation Assay is based on the reduction of tetrazolium salt WST-1 to a soluble formazan dye by electron transport across the plasma membrane of dividing cells. This kit will also allow investigators to screen drug candidates involved in cell cycle regulation.
TBARS Assay Kit Reference: 10009055-96 Oxidative stress in the cellular environment results in the formation of highly reactive and unstable lipid hydroperoxides. Decomposition of the unstable peroxides derived from PUFAs results in the formation of malondialdehyde (MDA), which can be quantified colorimetrically following its controlled reaction with thiobarbituric acid. The measurement of these TBARS is a well-established method for screening and monitoring lipid peroxidation. Cayman’s TBARS Assay provides a simple, reproducible, and standardized tool for assaying lipid peroxidation in plasma, serum, urine, tissue homogenates and cell lysates.
Caspase-3/7 Fluorescence Assay Kit Reference: 10009135-96 Cayman’s Caspase-3/7 Fluorescence Assay Kit employs a specific substrate, N-Ac-DEVD-N’-MC-R110, which upon cleavage by active caspase-3 or caspase-7, generates a highly fluorescent product that can be measured using excitation and emission wavelengths of 485 and 535 nm, respectively. The kit is simple to use and can be easily adapted to high throughput screening for therapeutic compounds regulating activation of caspase-3 or -7. Active caspase-3 is included in the kit for use as a positive control. The caspase-3/7 inhibitor, N-Ac-DEVD-CHO, is also included in the kit for verifying the specificity of the substrate.
JC-1 Mitochondrial Membrane Potential Assay Kit Reference: 10009172-100 Mitochondrial membrane potential, (ΔψM), is an important parameter of mitochondrial function used as an indicator of cell health. JC-1 is a lipophilic, cationic dye that can selectively enter into mitochondria and reversibly change color from green to red as the membrane potential increases. In healthy cells with high mitochondrial (ΔψM), JC-1 spontaneously forms complexes known as J-aggregates with intense red fluorescence. On the other hand, in apoptotic or unhealthy cells with low (ΔψM), JC-1 remains in the monomeric form, which shows only green fluorescence. Cayman’s JC-1 Mitochondrial Membrane Potential Assay Kit provides all the necessary reagents, as well as complete instructions, for analysis of mitochondrial integrity in whole cells. The assay is applicable to fluorescence microscopy and fluorescent plate reader. For flow cytometry, Cayman’s JC-1 Mitochondrial Membrane Potential Flow Cytometry Assay Kit (Item No. 701560) is recommended.
Malachite Green Phosphate Assay Kit Reference: 10009325-2 Cayman’s Malachite Green Phosphate Assay provides a fast, reproducible, colorimetric method for measuring inorganic free phosphate in aqueous solutions. The assay method is based on the formation of a complex between malachite green molybdate and free orthophosphate that absorbs at 620-640 nm. Applications for this assay include quantification of phosphorylation and phosphate release from protein phosphatase substrates. This assay is a reliable and suitable means of detecting and quantifying minimal amounts of inorganic free phosphate and is amenable to high-throughput screening applications. The assay is formatted to a 96-well format, but could easily be modified for use in 384-well or cuvette-based assays.
Cell Cycle Phase Determination Kit Reference: 10009349-100 Cayman’s Cell Cycle Phase Determination provides an easy to use tool for studying the induction and inhibition of cell cycle progression in any cell suspension sample. The assay involves the fixation and permeabilization of the cells of interest, making possible the staining of DNA within intact cells by propidium iodide. This kit will allow the investigator to determine the percentage of cells in a given sample that are within G1/G0, G2, or S phase at the time of fixation, as well as to quantify cells in the sub-G1 phase prior to apoptosis by FC.
MTT Cell Proliferation Assay Kit Reference: 10009365-480 Cayman’s MTT Proliferation Assay provides an easy to use tool for studying the induction and inhibition of cell proliferation in any in vitro model. This kit will also allow investigators to screen drug candidates involved in cell cycle regulation. In this assay, MTT is taken up by cells and then reduced to formazan by intracellular NAD(P)H-oxidoreductases.
Glucose Colorimetric Assay Kit Reference: 10009582-2 Cayman’s Glucose Assay provides a simple, reproducible, and sensitive tool for assaying glucose in plasma, serum, and urine. The glucose assay uses the glucose oxidase-peroxide reaction for the determination of glucose concentrations. In this assay, glucose is oxidized to d-gluconolactone with concomitant reduction of the flavin adenine dinucleotide (FAD)-dependent enzyme glucose oxidase. The reduced form of glucose oxidase is regenerated to its oxidized form by molecular oxygen to produce hydrogen peroxide. Finally, with horseradish peroxidase as a catalyst, hydrogen peroxide reacts with 3,5-dichloro-2-hydroxybenzenesulfonic acid and 4-aminoantipyrine to generate a pink dye with an optimal absorption at 514 nm.
Cholesterol Cell-Based Detection Assay Kit Reference: 10009779-2 The mechanism for the movement of cholesterol from intracellular sites to their ultimate cellular destination is an unresolved question of fundamental importance to cell biology and medicine. Thus, defining mechanisms of intracellular cholesterol transport and identifying the cellular factors involved are therefore of great interest. Cayman’s Cholesterol Cell-Based Detection Assay includes filipin III, fixative, and wash buffer in a ready to use format. It provides a simple fluorometric method to study mechanisms and biological factors that regulate cholesterol metabolism or movement within cells. A cholesterol trafficking inhibitor, U-18666A, is included as a positive control.
CREB (Phospho-Ser133) Transcription Factor Assay Kit Reference: 10009846-96 CREB (cAMP-response-element-binding protein) is a transcription factor that binds to cAMP-responsive element (CRE) promoter sites to regulate the transcription of numerous genes involved in metabolic regulation, depression, long term memory, and other physiological processes. Phosphorylation on serine 133 (Ser133) activates CREB to induce transcription of target genes. Diverse stimuli such as growth factors, neurotransmitters, hypoxia, growth factors, UV light, survival signals, and stress signals are some of the known activators of CREB. Cayman’s CREB (Phospho-Ser133) Transcription Factor Assay is a non-radioactive, sensitive method for detecting CREB DNA binding activity in nuclear extracts or whole cell lysates.