Flowcytometry TLR2 (CD282)

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 CD252 (OX40L) Antibody to perform Flow cytometry Anti-bodies Mouse - CD252/OX40L

Get tips on using PE/Cyanine7 anti-mouse CD252 (OX40L) Antibody to perform Flow cytometry Anti-bodies Mouse - CD252/OX40L

Get tips on using Biotin Rat Anti-Mouse OX40 Ligand (CD252) to perform Flow cytometry Anti-bodies Mouse - CD252/OX40L

Get tips on using FITC Mouse Anti-Human CD22 to perform Flow cytometry Anti-bodies Human - CD22

Get tips on using Purified Mouse Anti-Human CD22 to perform Flow cytometry Anti-bodies Human - CD22

Get tips on using BUV737 Mouse Anti-Human CD28 to perform Flow cytometry Anti-bodies Human - CD28

Get tips on using PE Mouse Anti-Human CD28 to perform Flow cytometry Anti-bodies Human - CD28

Get tips on using CD22-PE, SJ10.1H11, 2 mL, ASR to perform Flow cytometry Anti-bodies Human - CD22

Get tips on using PE-Cy™7 Mouse Anti-Human CD28 to perform Flow cytometry Anti-bodies Human - CD28