ChIP H3K27me3 Mouse Bovine Agarose

Get tips on using Mouse Myeloperoxidase DuoSet ELISA to perform ELISA Mouse - MPO

Get tips on using Mouse Leptin ELISA Kit to perform ELISA Mouse - Leptin

Get tips on using Mouse Decorin ELISA Kit to perform ELISA Mouse - Decorin

Get tips on using Mouse Osteopontin/OPN Antibody to perform Immunohistochemistry Mouse - Spp1/OPN

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 has 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 the efficacy of transduction and shRNA on its target site.

Get tips on using APC anti-mouse CD206 (MMR) Antibody to perform Flow cytometry Anti-bodies Mouse - CD206

Get tips on using Mouse Integrin alpha E/CD103 Antibody to perform Flow cytometry Anti-bodies Mouse - CD103

Get tips on using PE/Cyanine7 anti-mouse CD14 Antibody to perform Flow cytometry Anti-bodies Mouse - CD14

Get tips on using Mouse CD200 R3 PE-conjugated Antibody to perform Flow cytometry Anti-bodies Mouse - CD200

Get tips on using siGENOME Mouse Alox12 siRNA to perform siRNA / miRNA gene silencing Mouse - B16-F10 12-Lox/ALOX12