Protein Expression Eukaryotic cells BHK cells

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.

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - BHK-21

Get tips on using pPIC-xyn11B to perform Protein Expression Eukaryotic cells - P. pastoris Penicillium oxalicum xyn11B

Get tips on using pPIC-xyn11A to perform Protein Expression Eukaryotic cells - P. pastoris Penicillium oxalicum xyn11A

Get tips on using pPIC-xyn10B to perform Protein Expression Eukaryotic cells - P. pastoris Penicillium oxalicum xyn10B

Get tips on using pPIC-xyn10A to perform Protein Expression Eukaryotic cells - P. pastoris Penicillium oxalicum xyn10A

Get tips on using pPIC9K-anPhyA to perform Protein Expression Eukaryotic cells - P. pastoris Aspergillus niger PhyA

Get tips on using pPIC9/MoL to perform Protein Expression Eukaryotic cells - P. pastoris M. oleifera lectin