Immunohistochemistry 53BP2 phospho (ser-25) Rabbit IgG Human

Get tips on using peSpCas9(1.1)-2×sgRNA (empty, donor) to perform CRISPR Human - Repression SLC7A11

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Get tips on using TruSeq RNA Library Prep Kit v2 to perform RNA sequencing Human - MIA PaCa-2

Get tips on using TruSeq RNA Library Prep Kit v2 to perform RNA sequencing Human - iPSC-derived cardiomyocytes

Get tips on using TruSeq RNA Library Prep Kit v2 to perform RNA sequencing Human - MDA-MB-231

Get tips on using FITC Mouse Anti-Ki-67 Set to perform Flow cytometry Anti-bodies Human - Ki-67

I work with Human liver organoids and I would like to know the impact of FBS on the organoids. Since the composition of FBS is unknown, would you recommend any alternatives such as human platelet lysate?

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 SIHK1738 to perform siRNA / miRNA gene silencing Human - MDA-MB-231 PKN3

Get tips on using s4215 to perform siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b