Protein Expression Prokaryotic cells B. bifidum

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.

Get tips on using Strep-Tactin Superflow Plus (10 ml) to perform Protein tag Purification of Strep-tagged proteins

Get tips on using Ni-NTA Fast Start Kit (6) to perform Protein tag Purification of His-tagged proteins

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.

Get tips on using 2-D Quant Kit to perform Protein quantification Colorimetric method

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Mouse heart

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Mouse aorta