Wound healing assay cell type mouse

Get tips on using Atg3 Antibody to perform Autophagy assay cell type - KG1 cells

Get tips on using MouseTRAP™ (TRAcP 5b) ELISA to perform Acid phosphatase assay cell type - murine macrophage cells

Get tips on using PCNA Antibody (F-2) to perform Cell cytotoxicity / Proliferation assay cell type - oral squamous cell carcinoma

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 Dead Cell Apoptosis Kit with Annexin V Alexa Fluor™ 488 & Propidium Iodide (PI) to perform Apoptosis assay cell type - RPMI-8226

Get tips on using Dead Cell Apoptosis Kit with Annexin V Alexa Fluor™ 488 & Propidium Iodide (PI) to perform Apoptosis assay cell type - SH-SY5Y

Get tips on using In Vitro Angiogenesis Assay Kit to perform Angiogenesis assay mouse - MS1

Get tips on using Cytotoxicity Detection KitPLUS (LDH) to perform Cell cytotoxicity / Proliferation assay cell type - HT-22

Get tips on using LDH Cytotoxicity Detection Kit to perform Cell cytotoxicity / Proliferation assay cell type - 3T3-L1

Get tips on using Beclin-1 (D40C5) Rabbit mAb to perform Cell cytotoxicity / Proliferation assay cell type - K562