Cell cycle assay mouse

Get tips on using Host Cell Residual DNA contamination LANCE Ultra TR-FRET Detection Kit, 500 Assay Points to perform Cell Culture Contamination Detection Kit Bacteria

Get tips on using Live/Dead Cell Double Staining Kit to perform Live / Dead assay mammalian cells - L29 mouse fibroblast

Get tips on using Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO) to perform Autophagy assay cell type - THP 1

Get tips on using Purified Mouse Anti-p62 Ick ligand Clone 3/P62 LCK LIGAND (RUO) to perform Autophagy assay cell type - SH-SY5Y

Short hairpin or small hairpin RNA (shRNA) is artificial RNA, which has a hairpin loop structure, and uses inherent microRNA (miRNA) machinery to silence target gene expression. This is called RNA interference (RNAi). These can be delivered via plasmids or viral/bacterial vectors. Challenges in shRNA-mediated gene silencing include: 1. Off-target silencing, 2. Packaging shRNA encoding lentivirus, and 3. Stable transduction in cells. RNAi have been designed to have anywhere from 19-27 bs, but the most effective design has 19 bp. In case commercial shRNAs are not available, potential target sites can be chosen within exon, 5’- or 3’ UTR, depending on which splice variants of the gene are desired. One should use the latest algorithms and choose at least two different sequences, targeting different regions, in order to have confidence in overcoming off-target effects. A BLAST search after selecting potential design will eliminate potential off-target sequences. For the second challenge, sequencing the vector using primers for either strand (50-100 bp upstream) is suggested, along with using enzymatic digestion on agarose gel for the vector. Next, once the shRNA-containing vector is packaged in a virus, it is important to check the viral titer before transduction. Finally, using a marker in the lentiviral vector (fluorescent protein or antibiotic resistance), along with qPCR for target gene expression can help in determining efficacy of transduction and shRNA on its target site.

Get tips on using In Vitro Angiogenesis Assay Kit to perform Angiogenesis assay mouse - MS1

Get tips on using LIVE/DEAD Fixable Violet Dead Cell Stain Kit to perform Live / Dead assay mammalian cells - mouse splenocytes

HeLa cells I used are passage 40. Are these cells good to perform Cell cycle analysis on?

Get tips on using LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit to perform Live / Dead assay mammalian cells - mouse keratinocytes

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse cardiac fibroblasts