Protein isolation Mammalian cells

Get tips on using pET-21b(+)/Pro j 1 to perform Protein Expression Prokaryotic cells - E. coli Pro J 1

Get tips on using Quick-RNA™ MiniPrep Plus to perform RNA isolation / purification Cells - primary human epithelial cells

Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Cells - Primary cells HUVEC

Get tips on using His-Strep pQE-TriSystem Vector Set to perform Protein Expression Prokaryotic cells - E. coli Integrin αV

Get tips on using pFastBac1- B/Brisbane/60/2008-NP to perform Protein Expression Eukaryotic cells - S. frugiperda Influenza NP

Get tips on using pcDNA™3.1D/V5-His TOPO®-hsEH to perform Protein Expression Eukaryotic cells - HEK293 hsEH

The RNA interference (RNAi) is used to inhibit gene expression or translation, by neutralizing targeted mRNA molecules. Two types of RNA molecules such as microRNA (miRNA) and small interfering RNA (siRNA) play a central role in RNAi. Few points have to considered to increase the transfection efficiency of siRNA. Always use healthy, actively dividing cells to maximize transfection efficiency. The confluency of cells should be between 50-70%. Always use the most appropriate siRNA concentration to avoid off-target effects and unwanted toxic side effects. Positive and negative controls should be used for each and every experiment to determine transfection efficiency.

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.