siRNA / miRNA gene silencing Human HeLa EPAS-1

Get tips on using AllPrep DNA/RNA/miRNA Universal Kit to perform RNA isolation / purification Tissue - human adipose tissue

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells

Get tips on using Ki-67 Antigen, Clone MIB-1 to perform Immunohistochemistry Ki67 - Rabbit Mouse / Human -NA-

Get tips on using Notch 1 Antibody (A-8): sc-376403 to perform Immunohistochemistry Human - Notch1

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 miRNA Complete Labeling and Hyb Kit to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Cyanine 3-pCp

Get tips on using Beclin-1 Antibody to perform Autophagy assay cell type - THP 1

Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

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.