Flowcytometry CD14 Mouse / IgG2a Human

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using PE anti-human CD111 (Nectin-1) Antibody to perform Flow cytometry Anti-bodies Human - CD111/Nectin-1

Get tips on using PE anti-human CD126 (IL-6Rα) Antibody to perform Flow cytometry Anti-bodies Human - CD126/IL-6Ralpha

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 Monoclonal Anti-Proliferating Cell Nuclear Antigen antibody produced in mouse to perform Western blotting PCNA

Get tips on using PerCP/Cyanine5.5 anti-human CD127 (IL-7Rα) Antibody to perform Flow cytometry Anti-bodies Human - CD127

Get tips on using Monoclonal Mouse Anti-Villin (Autostainer Link 48) Clone 1D2 C3p to perform DNA Damage Assay U266 -

Get tips on using Mouse Monoclonal Antibody to Lamin A/C Cat# MCA-4C4 to perform Western blotting Lamin A/C

Get tips on using Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA Kit to perform ELISA Rat - TGF-beta 1