NETosis Assay Kit Reference: 601010-1 Cayman's NETosis Assay Kit provides a simple and fast method for studying the process of NETosis ex vivo. Notably, Cayman's NETosis Assay Kit does not depend upon the DNA component of neutrophil extracellular traps (NETs), as DNA release can occur independently of NETosis. In this kit, primary neutrophils are stimulated to release NETs with either PMA or a calcium ionophore (both included). Unbound neutrophil elastase is washed away following NET generation. Following digest of NET DNA by S7 nuclease, the supernatant containing neutrophil elastase is added to a substrate, which is selectively cleaved by elastase to yield a 4-nitroaniline product that adsorbs light at 405 nm. Enough reagents are provided to test 24 sample conditions for NET production, with analysis in duplicate.
Oct4 Transcription Factor Assay Kit Reference: 601080-96 Oct4 is an essential regulator of embryonic stem cell pluripotency and used as a marker for undifferentiated cells. It also plays a role in gene expression, DNA repair and/or replication, signal transduction and tumorigenesis. Cayman’s Oct4 Transcription Factor Assay is a non-radioactive, sensitive method for detecting specific transcription factor-DNA binding activity in nuclear extracts. A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double-stranded DNA (dsDNA) sequence containing the Oct4 response element is immobilized onto the wells of a 96-well plate. Oct4, contained in a nuclear extract, binds specifically to the Oct4 response element. Oct4 is detected by the addition of a specific primary antibody directed against Oct4. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.
Protein Synthesis Assay Kit Reference: 601100-96 Cayman’s Protein Synthesis Assay Kit is a non-radioactive method for measuring protein synthesis that employs the cell-permeable, alkyne-containing, puromycin analog O-Propargyl-puromycin (OPP). Upon application to cells, the OPP probe incorporates into the C-terminus of translating polypeptide chains, thereby stopping translation. These truncated C-terminal alkyne-labeled proteins are then subsequently detected via copper-catalyzed click chemistry using 5 FAM-Azide. The reagents provided in the kit are sufficient to run 20 samples when using flow cytometry or 100 samples when using a 96-well plate format.