Get tips on using Pierce™ Magnetic ChIP Kit to perform ChIP Human - Fibroblast cell lines
Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Mouse - 3T3-L1 cells
Get tips on using Pierce™ Magnetic ChIP Kit to perform ChIP Mouse - 3T3-L1 cells
Get tips on using NF-κB p65 (D14E12) XP® Rabbit mAb #8242 to perform Immunohistochemistry Mouse - NFκB / p65
Get tips on using OSTEOPONTIN (O-17) ANTI-HUMAN RABBIT IGG AFFINITY PURIFY to perform Immunohistochemistry Mouse - Spp1/OPN
Get tips on using Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb #2078 to perform Western blotting IKK Alpha
Get tips on using Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb #4806 to perform Western blotting NF-kB
Get tips on using High-Sensitivity ChIP Kit (ab185913) to perform ChIP Mouse - MLL-AF9/NrasG12D AML
Get tips on using Anti-PI3-kinase p85-α antibody produced in rabbit to perform Autophagy assay cell type - HepG2
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.
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