Protein Expression Eukaryotic cells HeLa R19

Get tips on using pET-21b(+)/Pro j 1 to perform Protein Expression Prokaryotic cells - E. coli Pro J 1

Get tips on using His-Strep pQE-TriSystem Vector Set to perform Protein Expression Prokaryotic cells - E. coli Integrin αV

Get tips on using Viromer® RED to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using Guava Nexin Annexin V Assay to perform Apoptosis assay cell type - HeLa cells

Get tips on using pPZP-RCS2-ocs-nptII to perform Protein Expression Prokaryotic cells - A. tumefaciens α-amylase, amylopullulanase, and glucoamylase

Get tips on using pET30 Ek/LIC-Colα3(IV) NC1 to perform Protein Expression Prokaryotic cells - E. coli Colα3(IV) NC1

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized HeLa, CaSki and C33A (Cervical cancer cells)

Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Cells - Immortalized cell lines HeLa

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

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.