Protein Expression Eukaryotic cells HeLa R19

Get tips on using Anti-Clara Cell Secretory Protein Antibody to perform Flow cytometry Anti-bodies Mouse - CCSP

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

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 LowCell# ChIP kit protein A x16 to perform ChIP Human - HepG2

Get tips on using heat shock protein family A (Hsp70) member 5 to perform siRNA / miRNA gene silencing Human - PC3 (human prostate cancer cell line) HSPA5 (GRP78)

Get tips on using EpiTect MSP Kit to perform DNA methylation profiling Gene specific profiling - HeLa MEG3

Get tips on using siGENOME Human PAK1 siRNA to perform siRNA / miRNA gene silencing Human - HeLa PAK1

Get tips on using EZ‐ChIP™ Assay Kit (Cat#17–371) to perform ChIP Human - HeLa

Get tips on using Mouse C Reactive Protein ELISA Kit (PTX1) (ab157712) to perform ELISA Mouse - C-Reactive Protein/CRP

Get tips on using Rat Bone Morphogenetic Protein 2 ELISA to perform ELISA Rat - BMP-2