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16:57, 2 March 2022 Admin talk contribs uploaded File:50mm 2well Long.png
16:56, 2 March 2022 Admin talk contribs created page File:50mm 2well.png
16:56, 2 March 2022 Admin talk contribs uploaded File:50mm 2well.png
16:55, 2 March 2022 Admin talk contribs created page File:50mm 1well.png
16:55, 2 March 2022 Admin talk contribs uploaded File:50mm 1well.png
13:44, 27 February 2022 Dicksonb talk contribs created page File:96 well plate for wiki (1).png
13:44, 27 February 2022 Dicksonb talk contribs uploaded File:96 well plate for wiki (1).png
17:13, 18 January 2022 Admin talk contribs created page Nucleofector (Created page with "Protocols and settings for the nucleofector. ==Reusing Cuvettes== Nucleofector cuvettes can be reused several times, allowing for far more transfections to be conducted than the kits claim. Reuse is limited to 5-8 transfections, and it is a good idea to try and use the same cuvette for the same plasmids to limit the risk of cross-contamination. Decreasing transfection efficiency, or sparking during electroporation, are signs that the cuvette is no longer usable. ===Cle...") Tag: Visual edit
18:46, 14 January 2022 Dicksonb talk contribs created page File:Bradford plate layout.png
18:46, 14 January 2022 Dicksonb talk contribs uploaded File:Bradford plate layout.png
01:34, 14 January 2022 Dicksonb talk contribs created page Traction Force Microscopy (In progress) Tag: Visual edit
15:51, 13 January 2022 Dicksonb talk contribs created page Bradford Assay (The Bradford assay is used to determine the concentration of total protein within a sample relative to a standard curve.)
17:16, 11 January 2022 Akipp4 talk contribs created page Transduction of THP-1s (Created page with "'''Transduction Reaction''' * The volume of the reaction is based on the amount of virus available, the number of cells and the MOI being used * Prepare the mixture as follows: ** X uL of concentrated virus ** X uL of SF Macrophage Media (chose this volume based on the vessel the cells will be grown in ** 1000X polybrene '''Protocol''' # Count cells, add cells to appropriate size tube, spin down at 400 x G for 5 minutes # Re-suspend cells in the transduct...") Tag: Visual edit
15:17, 25 November 2021 Akipp4 talk contribs created page Titering Pseudo-typed Lentiviruses (Created page with "- for titering one “batch” of collected virus '''Cell Seeding''' 1. Seed 6 wells of a 12 well plate with 30 000 – 50 000 HeLas per well on coverslips '''Transduction''' 1. Prepare “transduction media” by mixing 6 µL of polybrene (1000X) in 6 mL of DMEM+10% FBS 2. Remove media from the plate and replace with 900 µL of transduction media in each well 3. Begin the serial dilution by adding 100 µL of virus to the first we...") Tag: Visual edit
18:50, 18 November 2021 Admin talk contribs created page Immuno-FISH (Created page with "This is a protocol for performing Fluorescence In-Situ Hybridization alongside immunostained cells. The protocol assumes that the cells are already grown on a coverslip, treat...") Tag: Visual edit
18:26, 17 November 2021 Admin talk contribs created page File:FRET equasion.png
18:26, 17 November 2021 Admin talk contribs uploaded File:FRET equasion.png
18:21, 17 November 2021 Admin talk contribs created page FRET in FIJI (Created page with "There are two methods that can be easily implemented in FIJI and on our microscope to calculate FRET ratios in fixed or live cell images. The methods, and FIJI scripts to auto...") Tag: Visual edit
14:34, 16 November 2021 Admin talk contribs created page Nitrogen Cavitation (Created page with "Nitrogen caviatation is a gentle way of lysing cells, and can be used to recover intact organelles, efferosomes, phagosomes, etc. We have two protocols, optimized for differen...") Tag: Visual edit
14:47, 15 November 2021 User account Dicksonb talk contribs was created by Admin talk contribs
13:17, 13 September 2021 User account Akipp4 talk contribs was created by Admin talk contribs
20:53, 24 March 2021 Kwodz talk contribs created page Preparation of Digestion-Tracking Bacteria (Created page with "1. Measure out 4 mL of LB media and add kanamycin to a final concentration of 50 µg/mL. 2. Inoculate with the Bacteria Digest 1 (Clone RC-094) bacteria, and grow overnight s...")
12:46, 4 February 2021 Admin talk contribs changed group membership for Bheit from interface administrator and administrator to interface administrator, administrator and bureaucrat
12:37, 4 February 2021 User account Alac2 talk contribs was created by Admin talk contribs
12:36, 4 February 2021 User account Kwodz talk contribs was created by Admin talk contribs
12:35, 4 February 2021 Admin talk contribs changed group membership for Bheit from (none) to administrator and interface administrator
12:33, 4 February 2021 User account Bheit talk contribs was created by Admin talk contribs
20:14, 1 February 2021 Admin talk contribs created page File:PnW.png
20:14, 1 February 2021 Admin talk contribs uploaded File:PnW.png
19:58, 1 February 2021 Admin talk contribs created page Lipisome and Lipid-Coated Beads (Created page with "= General Notes = *All buffers should be filtered through a 0.2um filter to remove any particulates before use *Avoid plastics until beads/lipisomes are generated, as lipids...")
19:58, 1 February 2021 Admin talk contribs created page Lipid Extraction from Cells (Created page with "= Protocol = ===== Initial extraction: ===== #Wash cells 3X with PBS to remove serum. #Remove cells by scraping cells into PBS. #Pellet cells in ultra centrifuge; max spee...")
19:58, 1 February 2021 Admin talk contribs created page Lipid Coated Bead Preparation (Created page with "= Protocol = ===== Lipid Preparation: ===== #In a glass container, and using lipids dissolved in chloroform, mix desired lipids to a final quantity of ~4mmol total lipid. #...")
19:58, 1 February 2021 Admin talk contribs created page Asymmetric liposomes (Created page with "= Protocol = ===== Outer Bilayer Lipid Preparation: ===== #Measure out the lipid mixtures into sialinized glass vials. Evaporate the chloroform using nitrogen or argon....")
19:57, 1 February 2021 Admin talk contribs created page 3D Printed PDMS Chambers (Created page with "== Introduction == Highly detailed imaging chambers can be constructed by 3D printing a mould using DLP (resin) printing, which is then used to cast a PDMS chamber that can be...")
19:57, 1 February 2021 Admin talk contribs created page Live Cell FRET (Created page with "Our microscope is capable of FRET measurements using conventional CFP/YFP pairs and their more modern derivatives. We recommend using [https://www.fpbase.org/protein/mturquois...")
19:57, 1 February 2021 Admin talk contribs created page Acid Washing Coverslips (Created page with "'''WARNING: These procedures involve the use of heated strong acids. Clearly label all containers with warning signs, and wear all appropriate safety equipment.''' Acid clean...")
19:56, 1 February 2021 Admin talk contribs created page Staining for GSD (Created page with "Ground State Depletion (GSD) microscopy detects fluorophore positions independently of intensity or staining density. As such, it is critical that p...")
19:56, 1 February 2021 Admin talk contribs created page Single Particle Tracking (Created page with "= Notes = *For imaging, use a space ship with the inner surface painted flat black *The dilution for the primary Fab' must be worked out for each antibody, and after each di...")
19:56, 1 February 2021 Admin talk contribs created page Inhibition of Focal Contact Signaling (Created page with "= Protocol = #Cool cells if needed, protocol also works at 37C #Wash 2X with PBS at required temperature #Cover cells with PEG media, incubate 10-20 min #Wash carefully wi...")
19:56, 1 February 2021 Admin talk contribs created page Immunostaining (Created page with "This procedure was optimized using the 4G10 (anti-phosphotyrosine) antibody, but should work well for many other antibodies. For a generic protocol, add an additional 20-60min...")
19:56, 1 February 2021 Admin talk contribs created page Competition of Charged Molecules with Lipophilic Cations (Created page with "= Protocol = #TF cells on coverslips with desired construct, and treat cells as required for the experiment. #Prepare 2x stocks of desired lipophilic compound (see table...")
19:55, 1 February 2021 Admin talk contribs created page Apoptosis Detection with AnnexinV and PI (Created page with "==Reagents== *Binding Buffer *AnnexinV-flourophore conjugate (e.g. ThermoFisher AnnexinV-488) *Propridium Iodide ==Protocol== ===Binding Buffer=== Prepare binding buffer, for...") Tag: Visual edit: Switched
19:54, 1 February 2021 Admin talk contribs created page G418 & Puromycin Kill Curves (Created page with "= G418 and Puromycin Kill Curves = G418 and puromycin can be used to generate stabally transfected cell lines. However, the concentratio of these antibiotics that are require...")
19:54, 1 February 2021 Admin talk contribs created page Cell-Type Specific Transfection Protocols (Created page with "This page contains the protocol of transfecting specific cell types. Unless otherwise noted, all transfections are for 12-well (22mm) plates. == RAW 264.7 Mur...")
19:54, 1 February 2021 Admin talk contribs created page Ablation of Recycling Endosomes (Created page with "From [http://www.ncbi.nlm.nih.gov/pubmed/15534004 Ang et al. 2004, JCB 167: 531] == Stock solutions == ===== Human Tfn-HRP ===== *0.5 mg/ml in H2O + 20% glycerol ===...")
19:53, 1 February 2021 Admin talk contribs created page Stripping & Reprobing Blots (Created page with "It is possible to strip the detection antibodies off of a western blot, and then re-probe that blot with a second set of antibodies. Every strip will reduce the amount o...")
19:53, 1 February 2021 Admin talk contribs created page Receptor Cross-linking and Activation (Created page with "Ligand-independent activation of most receptors can be achieved through antibody-mediated cross-linking. However, care must be taken, especially on cels of the immune sy...")
19:53, 1 February 2021 Admin talk contribs created page Coomassie Staining (Created page with "Thes procedures are adapted from the procedure at [http://www.nationaldiagnostics.com/article_info.php/articles_id/80 National Diagnostics]. = Method 1: Standard (sensitive)...")
19:53, 1 February 2021 Admin talk contribs created page Updated FPLC Size Exclusion Procedure (Created page with "==Important Notes == * All buffers, 20% ethanol and samples must be pre-filtered using a 0.2 μm filter * Install desired loading loop onto system before initiation, but do...")
19:53, 1 February 2021 Admin talk contribs created page Fab Purification by FPLC (Created page with "= Notes = *All buffers must be filtered or otherwise rendered particle-free. *pH 2-10 can be used *Sterile PBS can be used as a buffer. *A maximum of 2ml can be run throu...") Tag: Visual edit: Switched

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