Site Directed Mutagenesis (SDM) Human Point mutation MDA-MB-231

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.

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.

A key signature for necrotic cells is the permeabilization of the plasma membrane. Necrosis can be quantified by several cellular and biochemical assays. When studied minutely, it reveals the difficulty in confirmation in secondary induction of necrosis in apoptotic cells. Apoptotic cells are being analyzed to shift to necrotic status owing to membrane permeability at later stages, and thus, discrimination of two cell death becomes critical. Therefore, it is crucial to use a necrosis detection kit or a defined procedure to analyze this unprogrammed form of death in response to immense chemical and physical insults.

Get tips on using Hoechst 33342 to perform Necrosis MIA PaCa-2

Get tips on using BstNI NEB#R0168 to perform Restriction Enzymes BstNI / MvaI

Get tips on using Fluoro hROS to perform ROS assay cell type - MiaPaCa-2 pancreatic carcinoma

Get tips on using Annexin V-FITC Apoptosis Kit to perform Necrosis MIA PaCa-2

Get tips on using Apoptosis and Necrosis Quantification Kit to perform Necrosis MIA PaCa-2

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Cells - immortalized MIA PaCa-2

Get tips on using NucleoSpin® RNA to perform RNA isolation / purification Cells - immortalized MIA PaCa-2