Protein Expression Eukaryotic cells Y. lipolytica

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - BHK-21

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.

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - HepG2

Get tips on using Mammalian Cell Lysis kit to perform Protein isolation Mammalian cells - STTG1

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Mouse Epididymal fat

Get tips on using RIPA Buffer (10X) to perform Protein isolation Mammalian cells - Rat_Renal tissue

Get tips on using TRI Reagent® to perform Protein isolation Mammalian cells - Mouse_Brown fat

Get tips on using RIPA Lysis Buffer, 10X to perform Protein isolation Mammalian cells - HaCaT