siRNA / miRNA gene silencing Human Human ovarian carcinoma cell (OV2008) Yap Gene

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Get tips on using β-Galactosidase Reporter Gene Staining Kit to perform Reporter gene assay β-galactosidase substrates - mouse embryo tissue

Products Sigma-Aldrich β-Galactosidase Reporter Gene Staining Kit

Get tips on using FITC Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - OV2008

Products BD Biosciences FITC Annexin V Apoptosis Detection Kit I

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Cellular assays Autophagy assay cell type MT-2 (human T cell leukaemia)

Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform Microarray Gene expression arrays - Rhesus monkey brain tissue Biotin

Products Thermo Fisher Scientific GeneChip® Human Genome U133 Plus 2.0 Array

Get tips on using CpGenome Universal DNA Modification Kit to perform DNA methylation profiling Whole genome profiling - OVCAR-3 human ovarian cancer

Products Millipore CpGenome Universal DNA Modification Kit

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

Products Cell Biolabs OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence)

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Cellular assays TUNEL assay cell type A127, U87MG, U251MG, T98G human glioblastoma cells

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

DNA Microarray Gene expression arrays Mouse Cyanine-CTP

RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.

RNA RNA sequencing Human HT-1376 (urinary bladder cell line)

Get tips on using APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU to perform TUNEL assay cell type - A549, NCI-H460, H1299 human alveolar carcinoma

Products Thermo Fisher Scientific APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU

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