Wound healing assay cell type

Flow cytometry is an immunophenotyping technique whereby sing cell suspensions are stained for either cell surface markers or intracellular proteins by fluorescently-labelled antibodies and analyzed with a flow cytometer, where fluorescently-labelled molecules are excited by the laser to emit light at varying wavelengths, which is then detected by the instrument. There are several key criteria which are required to be kept in mind while designing a flow experiment- 1. Antibody titration (optimal dilution of antibodies should be calculated in order to avoid over- or under- saturated signals for proper detection of surface and intracellular markers), 2. Precision (3 or more replicates of the sample should be used per experiment), 3. Specificity (proper isotype controls should be included in the experiment), 4. Day-to-day variability (experiments should be repeated 3 or more times to ensure consistency and avoid variability due to flow cytometer settings), 5. Antibody interaction (Fluorescence minus one or FMO should be used, which is the comparison of signals from panel minus one antibody vs. the full panel), and 6. Antibody stability (fluorescently-labelled antibodies should be stored at 4C).

Generally isolating RNA from Gram-negative bacteria is easy, however keeping your working environment clean and RNase free (use RNase inhibitor) is essential. Some common points to keep in mind: a) Use fresh samples for isolation or store them by freezing in RNA stabilizing buffer until use. b) Choose the bacterial input amounts carefully, to ensure buffer volumes are adequate and not to overload the columns.

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram negative Salmonella typhi

Get tips on using Qproteome Bacterial Protein Prep Kit to perform Protein isolation Bacteria - Salmonella typhi

Get tips on using Qproteome Bacterial Protein Prep Kit to perform Protein isolation Bacteria - Salmonella typhimurium

Get tips on using TRI Reagent™ Solution to perform RNA isolation / purification Bacteria - Gram negative Salmonella typhi

Get tips on using mericon DNA Bacteria Kit (100) to perform DNA isolation / purification Bacteria - Gram negative Salmonella typhi

Get tips on using RNeasy Protect Bacteria Mini Kit to perform RNA isolation / purification Bacteria - Gram negative Salmonella typhi

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - Human eutopic endometrial stromal cells

DNA ladder is typically used as a reference to estimate the size of unknown DNA samples that are separated based on their mobility in an electrical field. The critical points for running a DNA ladder are compatibility with running buffer, agarose gel percentage, and choosing the correct range of DNA ladder for sizing DNA molecules.