Whether your team is working in Miro or Mural, you can port your work directly over using InVision. I would appreciate if anyone shares it's experience in pulling down of endogenous proteins especially Miro proteins a have any recommendations for me to solve this problem and to improve my work. Seamlessly connect to the tools you use every day. All the attempt I made haven't been successful in terms of pulling down the interactor proteins beside the main protein so far. With Miro’s whiteboard mobile app, you can: Scan paper post-it notes and convert them into editable digital notes. With over 200+ pre-made templates, a drag-and-drop interface, and no limit on collaborators, working on our whiteboard is fast and fun. For both protocol I add protease inhibitor when I want to do CoIP. Miro’s whiteboard app allows you to create anytime, anywhere. The lysis buffer has 10mM Tris-HCl pH7.4, 150mM NaCl, 1.5mM MgCl2, 5m EDTA, 1% Triton, 10% glycerol, 1 mM NaF, 50mM Na2H2P2O7 and the washing buffer has 10mM TrisHCl pH 7.5, 500mM NaCl, 1% Triton, 0.5% NP-40, 1mM MgCl2, 1mM EDTA, 0.5mM EGTA. I wash the beads three times with low salt(150mM) and last wash I increase the salt concentration to 500mMthe The second protocol comes from a colleague and he could pull down overexpressed Miro1 and Miro2 together in HEK cells and I use Chromoteck Agarose beads to pull down Miro proteins. Aptly titled Le Marechal des Logis (The Sergeant) a badge of accolades is present in vibrant primary colors, just under an eye which stares out intimidatingly. Connect existing ways of working to Miro with 100+ apps and integrations like Zoom, Slack, Google Drive, and Sketch. It's lysis buffer has 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA, 0.5 %, Nonidet™ P40 Substitute, 0.09 % sodium azide and the washing buffer has 10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.05 % Nonidet™ P40, Substitute, 0.5 mM EDTA, 0.018 % sodium azide. A powerful black figure stares out from the composition emoting authority and confidence. The first one is from Chromoteck and I used magnetic beads to pull down the protein. When I use each of the MYC or GFP beads I can pull down the tagged protein easily with high efficiency but I can not detect other protein-interactor by western blotting. I made double tagged Miro1-GFP and Miro2-MYC stable HeLa cell line and I use MYC/GFP Agarose or Agarose magnetic beads to pull down those proteins together. We suppose that Miro proteins interact with each other and I want to see If I can pull them down together or not. ![]() ![]() They are mitochondrial protein and isoform of each other. I'm trying to CoIP endogenous Miro1 and 2 proteins in HeLa cells.
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