In this protocol, we describe a method to isolate such known (and unknown) protein complexes bound to a nuclear noncoding RNA.doi:10.1007/978-1-4939-6716-2_9Ranveer Singh JayaniAmanjot SinghDimple Notani
Protein–protein and protein–DNA isolation His-tagged protein–protein pull downs. 10 μM His-tagged bait protein was bound to His Mag Sepharose Ni beads (GE Healthcare) according to the manufacturer’s instructions. Samples were incubated in 50 mM Tris-HCl (pH 7.5), 500 mM NaCl an...
Nuclear protein was extracted from right gastrocnemius muscle using a NE-PER™Nuclear and Cytoplasmic Extraction Reagent Kit (Thermo Fisher Scientific, MA, USA), according to the kit protocol. Nuclear samples were analysed by SDS-PAGE and western blotting as described above. Histochemical analysis R...
The protocol involves (i) disruption of mycelial cells by blending in liquid nitrogen followed by suspension of cell contents in buffer containing high concentrations of protease and EDTA; (ii) deproteinization with phenol; (iii) cesium chloride/bisbenzimide density gradient centrifugation to separate...
TheEpiQuik™ Nuclear Extraction Kit simply applies our proprietary nuclear protein isolation buffers to cell/tissues. After treatment with pre-lysis and lysis buffers, the nuclear proteins are easily extracted for immediate use or storage at proper conditions. ...
The NE-PER Kit is a nuclear protein extraction method that involves simple, stepwise lysis of cells and centrifugal isolation of nuclear and cytoplasmic protein fractions. A benchtop microcentrifuge, tubes and pipettors are the only tools required. The NE-PER Reagents efficiently solubilize and separa...
Protein purification protocol GST-tagged proteins were produced using IPTG-inducible BL-21 bacteria. Cells were grown in LB media, pelleted and lysed by sonication in solution (pH = 7.5, Tris HCl 50mM, NaCl 100mM, DTT 1mM, EDTA-free protease Inhibitors, Lysozyme 100 ug/ml, DNase 5 ug/mL...
M-PER Mammalian Protein Extraction Reagent is not suitable for cell fractionation. We recommend using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Cat. No. 78833, 78835).Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center. Typical...
Isolation of complementary DNAs encoding a cerebellum-enriched nuclear factor I family that activates transcription from the mouse myelin basic protein pro... and Mikoshiba, K., Isolation of complementary DNAs encoding a cerebellum-enriched nuclear factor I family that activates transcription from the ...
Cells were fractionated with Cell Surface Protein Isolation Kit (#89881, Thermo Fisher Scientific) according to the manufacturer’s protocol. Membranes were incubated with actin (#3280, 1:5,000, Abcam), β-tubulin (#ab6046, 1:5,000 Abcam), EEA1 (#E7659, 1:1,000, Sigma-Aldrich), ...