ChIP acH3 Mouse Canine

Get tips on using Monoclonal Anti-MAP Kinase, Activated/monophosphorylated (Phosphothreonine ERK-1&2) antibody produced in mouse to perform Western blotting ERK

Get tips on using Anti-Ctip1/BCL-11A antibody [14B5] (ab19487) to perform ChIP Anti-bodies CtIP/BCL11A

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 Histone H3K9ac antibody to perform ChIP H3K9Ac - Sheep Rat -NA-

Get tips on using Histone H2A.X antibody (pAb) to perform ChIP Anti-bodies γH2AX

Get tips on using Histone H4K20me1 antibody (mAb) to perform ChIP Anti-bodies H4K20me1

Get tips on using Histone H3K36me3 antibody (mAb) to perform ChIP Anti-bodies H3K36me3

Get tips on using Histone H3K9me3 antibody (mAb) to perform ChIP Anti-bodies H3K9me3

Get tips on using Histone H3K4me3 antibody (mAb) to perform ChIP Anti-bodies H3K27me1