ChIP H3K27me3 Goat Mouse

Get tips on using GATA-1 shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 GATA‐1

Get tips on using GATA-4 Antibody (G-4): sc-25310 to perform Western blotting GATA-4

Get tips on using siGENOME Rat Gata1 (25172) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - C6 (rat glioma) Gata1

Get tips on using pET-28a-chBCO2 to perform Protein Expression Prokaryotic cells - E. coli chicken BCO2

Get tips on using TRI Reagent® to perform Protein isolation Mammalian cells - Mouse_Brown fat

Get tips on using Gateway™ pDONR™221 Vector to perform Protein expression and purification Yeast - Saccharomyces cerevisiae ΔHSPA5

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.

Get tips on using Mem-PER™ Plus Membrane Protein Extraction Kit to perform Protein isolation Mammalian cells - Mouse_Brown fat

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.