Get tips on using SuperFect Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human aortic smooth muscle cells (HOSMC)
Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)
Get tips on using Polyclonal Rabbit Anti-Human Myeloperoxidase (Dako Omnis) to perform Immunohistochemistry Mouse - MPO
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 Human MCP-1 / CCL2 PicoKine™ ELISA Kit to perform ELISA Human - MCP1
Get tips on using Human Von Willebrand Factor ELISA Kit (VWF) (ab108918) to perform ELISA Human - VWF-A2
Get tips on using Human Lipocalin-2/NGAL PicoKine™ ELISA Kit to perform ELISA Human - NGAL/LCN2
Get tips on using Human KIM1 / TIM-1 PicoKine™ ELISA Kit to perform ELISA Human - KIM-1
Get tips on using Human TIM-1/KIM-1/HAVCR DuoSet ELISA to perform ELISA Human - KIM-1
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