Protein Expression Prokaryotic cells S. acidocaldarius

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - HEK293T

Get tips on using QuantiPro™ BCA Assay Kit to perform Protein quantification Mammalian cells - L929

Get tips on using QuantiPro™ BCA Assay Kit to perform Protein quantification Mammalian cells - RAW264.7

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HUVEC

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HeLa

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - SK-N-BE(2)-C

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - SK-N-BE(2)-C

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that no responses other than those related to the signaling pathway of interest. This can be achieved by selecting a highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzyme such as luciferase.

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase