ChIP acH4 Bovine Mouse

Get tips on using RIP-Assay Kit for microRNA to perform ChIP Human - MCF-7

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Human - MCF-7

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Rat - Brain microvessels

Get tips on using truChIP Chromatin Shearing Kit with Formaldehyde to perform ChIP Human - T47D

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Human - MDA-MB-231

Get tips on using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 to perform ChIP Anti-bodies H3K9-Ac

Get tips on using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733 to perform ChIP Anti-bodies H3K27me3

Get tips on using Mono-Methyl-Histone H3 (Lys27) (D3R8N) Rabbit mAb #84932 to perform ChIP Anti-bodies H3K27me1

Get tips on using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725 to perform ChIP Anti-bodies H3K4me2

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.