ChIP acH4 Bovine Rat

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

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

Get tips on using Anti-CCSP - Rabbit to perform Flow cytometry Anti-bodies Mouse - CCSP

Get tips on using CD34 Monoclonal Antibody (RAM34), eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD34

Get tips on using Recombinant Anti-CD204/MSR1 Antibody (FITC), Rabbit Monoclonal to perform Flow cytometry Anti-bodies Mouse - CD204

Get tips on using Ras (D2C1) Rabbit mAb #8955 to perform Western blotting Ras

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Liver

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Circumvallate papillae

Get tips on using RIPA Buffer (10X) to perform Protein isolation Mammalian cells - Rat_Renal tissue

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.