RNA isolation / purification Tissue Human

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

Get tips on using QuantiTect Virus Kit to perform RNA quantification qPCR

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid-mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time-consuming, but provide a more permanent expression of RNAi in the cells and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines.

Get tips on using In Vitro ROS/RNS Assay to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

Get tips on using RNeasy PowerClean Pro Cleanup Kit (50) to perform Removal of contamination in RNA Melanin contamination

Get tips on using Rab 7 siRNA (h) to perform siRNA / miRNA gene silencing Human - BEAS-2B RAB7A

Get tips on using Rab 5C siRNA (h) to perform siRNA / miRNA gene silencing Human - BEAS-2B RAB5C

Get tips on using 53BP1 Antibody (H-300) rabbit polyclonal to perform Immunohistochemistry 53BP1 - Rabbit IgG Human -NA-

Get tips on using Vimentin (D21H3) XP® Rabbit mAb to perform Immunohistochemistry Vimentin - Rabbit Human / mouse -NA-