siRNA / miRNA gene silencing Human Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1)

Get tips on using High Pure RNA Isolation Kit to perform RNA isolation / purification Cells - primary human dermal fibroblasts

Get tips on using AllPrep DNA/RNA Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes

Get tips on using Complete Kit for Human Whole Blood CD34+ Cells to perform Cell Isolation CD34+ cells

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Get tips on using Gibco DMEM/F-12, HEPES to perform 3D Cell Culture Media Human primary breast ephitelial cells-organoids

Get tips on using pSA-HVif-FabV to perform Protein Expression Prokaryotic cells - E. coli HIV-1 vif

Get tips on using pSA-HNef-FabV to perform Protein Expression Prokaryotic cells - E. coli HIV-1 nef

Get tips on using Mucin 1 Antibody (VU4H5): sc-7313 to perform Immunohistochemistry Human - Muc-1

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.