siRNA / miRNA gene silencing Human Caco-2

Get tips on using Monoclonal Anti-ATG12 antibody produced in mouse to perform Autophagy assay cell type - CaCo-2

Get tips on using VWR Life Science RIPA Lysis Buffer, Biotechnology Grade to perform Protein isolation Mammalian cells - Caco-2

Get tips on using LC3A/B (D3U4C) XP® Rabbit mAb #12741 to perform Autophagy assay cell type - CaCo-2

Get tips on using SLC20A2 Human Gene Knockout Kit to perform CRISPR Human - Repression SLC20A2

Get tips on using FOXA2 Human Gene Knockout Kit to perform CRISPR Human - Repression FOXA2

Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 GLUT1

Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 CD74

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 anti-p62 / SQSTM1 (C-terminus) guinea pig polyclonal, serum to perform Autophagy assay cell type - CaCo-2

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.