CRISPR Mouse Activation 3T3-L1

Get tips on using Xfect™ Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines 3T3-L1

Get tips on using GeneJuice® Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines 3T3-L1

Get tips on using MTT Cell Viability Assay Kit to perform Cell cytotoxicity / Proliferation assay cell type - 3T3-L1

Get tips on using Calcein AM Cell Viability Assay Kit to perform Live / Dead assay mammalian cells - 3T3-L1

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Human - Activation ERαY537S

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines 3T3-L1

Get tips on using PRO-PREP™ Protein Extraction Solution (C/T) to perform Protein isolation Mammalian cells - 3T3-L1

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - 3T3-L1

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.