Site Directed Mutagenesis (SDM) Human Point mutation BxPC-3

Get tips on using Apo-ONE® Homogeneous Caspase-3/7 Assay to perform Apoptosis assay cell type - SH-SY5Y

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Gene expression arrays - Mouse brain tissue Biotin

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray Gene expression arrays - Mouse dorsal skin Biotin

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray RNA amplification & Labeling - Mouse brain tissue Biotin

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.

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

Get tips on using Anti-PI 3 Kinase p85 alpha (phospho Y607) antibody to perform Autophagy assay cell type - HepG2

Get tips on using ON-TARGETplus Human GAPDH (2597) siRNA - Individual to perform siRNA / miRNA gene silencing Human - Calu-3 GAPDH