Protein expression and purification Mammalian cells HEK 293

Get tips on using PL_MmEro1l to perform Protein Expression Eukaryotic cells - CHO Ero1l

Get tips on using PL_MmBax to perform Protein Expression Eukaryotic cells - CHO Bax

Get tips on using PL_MmBak1 to perform Protein Expression Eukaryotic cells - CHO Bak1

Get tips on using PL_MmAtf4 to perform Protein Expression Eukaryotic cells - CHO Atf4

Get tips on using CpGfree to perform Protein Expression Eukaryotic cells - CHO EGFP

Get tips on using CpGrich to perform Protein Expression Eukaryotic cells - CHO EGFP

Get tips on using AllPrep RNA/Protein Kit to perform RNA isolation / purification Cells - primary human epidermal keratinocytes

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.

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 AcP(−)p6.9hEPO to perform Protein Expression Eukaryotic cells - S. frugiperda hEPO